Isolated human secreted proteins, nucleic acid molecules encoding human secreted proteins, and uses thereof

ABSTRACT

The present invention provides amino acid sequences of peptides that are encoded by genes within the human genome, the secreted peptides of the present invention. The present invention specifically provides isolated peptide and nucleic acid molecules, methods of identifying orthologs and paralogs of the secreted peptides, and methods of identifying modulators of the secreted peptides.

FIELD OF THE INVENTION

[0001] The present invention is in the field of secreted/extracellular matrix proteins that are related to the nephronectin subfamily, recombinant DNA molecules, and protein production. The present invention specifically provides a novel nephronectin protein splice form and nucleic acid molecules encoding the novel splice form, all of which are useful in the development of human therapeutics and diagnostic compositions and methods.

BACKGROUND OF THE INVENTION

[0002] Secreted Proteins Many human proteins serve as pharmaceutically active compounds. Several classes of human proteins that serve as such active compounds include hormones, cytokines, cell growth factors, and cell differentiation factors. Most proteins that can be used as a pharmaceutically active compound fall within the family of secreted proteins. It is, therefore, important in developing new pharmaceutical compounds to identify secreted proteins that can be tested for activity in a variety of animal models. The present invention advances the state of the art by providing many novel human secreted proteins.

[0003] Secreted proteins are generally produced within cells at rough endoplasmic reticulum, are then exported to the golgi complex, and then move to secretory vesicles or granules, where they are secreted to the exterior of the cell via exocytosis.

[0004] Secreted proteins are particularly useful as diagnostic markers. Many secreted proteins are found, and can easily be measured, in serum. For example, a ‘signal sequence trap’ technique can often be utilized because many secreted proteins, such as certain secretory breast cancer proteins, contain a molecular signal sequence for cellular export. Additionally, antibodies against particular secreted serum proteins can serve as potential diagnostic agents, such as for diagnosing cancer.

[0005] Secreted proteins play a critical role in a wide array of important biological processes in humans and have numerous utilities; several illustrative examples are discussed herein. For example, fibroblast secreted proteins participate in extracellular matrix formation. Extracellular matrix affects growth factor action, cell adhesion, and cell growth. Structural and quantitative characteristics of fibroblast secreted proteins are modified during the course of cellular aging and such aging related modifications may lead to increased inhibition of cell adhesion, inhibited cell stimulation by growth factors, and inhibited cell proliferative ability (Eleftheriou et al., Mutat Res 1991 March-November;256(2-6):127-38).

[0006] The secreted form of amyloid beta/A4 protein precursor (APP) functions as a growth and/or differentiation factor. The secreted form of APP can stimulate neurite extension of cultured neuroblastoma cells, presumably through binding to a cell surface receptor and thereby triggering intracellular transduction mechanisms. (Roch et al., Ann N Y Acad Sci Sep. 24, 1993 695:149-57). Secreted APPs modulate neuronal excitability, counteract effects of glutamate on growth cone behaviors, and increase synaptic complexity. The prominent effects of secreted APPs on synaptogenesis and neuronal survival suggest that secreted APPs play a major role in the process of natural cell death and, furthermore, may play a role in the development of a wide variety of neurological disorders, such as stroke, epilepsy, and Alzheimer's disease (Mattson et al., Perspect Dev Neurobiol 1998; 5(4):337-52).

[0007] Breast cancer cells secrete a 52K estrogen-regulated protein (see Rochefort et al., Ann N Y Acad Sci 1986;464:190-201). This secreted protein is therefore useful in breast cancer diagnosis.

[0008] Two secreted proteins released by platelets, platelet factor 4 (PF4) and beta-thromboglobulin (betaTG), are accurate indicators of platelet involvement in hemostasis and thrombosis and assays that measure these secreted proteins are useful for studying the pathogenesis and course of thromboembolic disorders (Kaplar, Adv Exp Med Biol 1978; 102:105-19).

[0009] Vascular endothelial growth factor (VEGF) is another example of a naturally secreted protein. VEGF binds to cell-surface heparan sulfates, is generated by hypoxic endothelial cells, reduces apoptosis, and binds to high-affinity receptors that are up-regulated by hypoxia (Asahara et al., Semin Interv Cardiol 1996 September; 1 (3):225-32).

[0010] Many critical components of the immune system are secreted proteins, such as antibodies, and many important functions of the immune system are dependent upon the action of secreted proteins. For example, Saxon et al., Biochem Soc Trans 1997 May;25(2):383-7, discusses secreted IgE proteins.

[0011] For a further review of secreted proteins, see Nilsen-Hamilton et al., Cell Biol Int Rep 1982 September;6(9):815-36.

[0012] Nephronectin

[0013] The novel human protein, and encoding transcript, provided by the present invention is a novel human nephronectin splice variant. Nephronectin is an extracellular matrix protein. The nephronectin gene is known in the art to encode several alternatively spliced transcripts, including at least two previously identified human transcripts. The splice form provided by the present invention is a novel human splice form previously uncharacterized in the art, and is supported by mouse sequences.

[0014] Previous studies in the mouse have indicated that nephronectin is a ligand that binds and modulates integrin alpha8beta1 function in the embryonic kidney (Schmidt et al., J Cell Biol Jul. 23, 2001;154(2):447-58). Mice lacking integrin alpha8beta1 display deficient epithelial-mesenchymal interactions, which are necessary for proper kidney organogenesis. Furthermore, because nephronectin is widely expressed outside the kidney, it has been suggested that nephronectin plays wider roles in development (Schmidt et al., J Cell Biol Jul. 23, 2001;154(2):447-58). Therefore, novel human nephronectin splice forms are particularly useful as therapeutic targets for treating developmental disorders.

[0015] Secreted/extracellular matrix proteins, particularly members of the nephronectin protein subfamily, are a major target for drug action and development. Accordingly, it is valuable to the field of pharmaceutical development to identify and characterize previously unknown members of this subfamily of secreted proteins. The present invention advances the state of the art by providing previously unidentified human secreted proteins that have homology to members of the nephronectin protein subfamily.

SUMMARY OF THE INVENTION

[0016] The present invention is based in part on the identification of amino acid sequences of a novel human nephronectin splice form, as well as allelic variants and other mammalian orthologs thereof These unique peptide sequences, and nucleic acid sequences that encode these peptides, can be used as models for the development of human therapeutic targets, aid in the identification of therapeutic proteins, and serve as targets for the development of human therapeutic agents that modulate nephronectin activity in cells and tissues that express nephronectin. Experimental data as provided in FIG. 1 indicates nephronectin expression in head/neck, nervous tumor, colon, breast, and placenta tissue.

DESCRIPTION OF THE FIGURE SHEETS

[0017]FIG. 1 provides the nucleotide sequence of a cDNA molecule or transcript sequence that encodes the secreted protein of the present invention. (SEQ ID NO:1) In addition, structure and functional information is provided, such as ATG start, stop and tissue distribution, where available, that allows one to readily determine specific uses of inventions based on this molecular sequence. Experimental data as provided in FIG. 1 indicates expression in head/neck, nervous tumor, colon, breast, and placenta tissue.

[0018]FIG. 2 provides the predicted amino acid sequence of the secreted protein of the present invention. (SEQ ID NO:2) In addition structure and functional information such as protein family, function, and modification sites is provided where available, allowing one to readily determine specific uses of inventions based on this molecular sequence.

[0019]FIG. 3 provides genomic sequences that span the gene encoding the secreted protein of the present invention. (SEQ ID NO:3) In addition structure and functional information, such as intron/exon structure, promoter location, etc., is provided where available, allowing one to readily determine specific uses of inventions based on this molecular sequence. As illustrated in FIG. 3, SNPs were identified at 24 different nucleotide positions in the nephronectin gene of the present invention.

DETAILED DESCRIPTION OF THE INVENTION

[0020] General Description

[0021] The present invention is based on the sequencing of the human genome. During the sequencing and assembly of the human genome, analysis of the sequence information revealed previously unidentified fragments of the human genome that encode peptides that share structural and/or sequence homology to protein/peptide/domains identified and characterized within the art as being a secreted/extracellular matrix protein or part of a secreted/extracellular matrix protein and are related to the nephronectin protein subfamily. Utilizing these sequences, additional genomic sequences were assembled and transcript and/or cDNA sequences were isolated and characterized. Based on this analysis, the present invention provides amino acid sequences of a novel human nephronectin splice form, nucleic acid sequences in the form of transcript sequences, cDNA sequences and/or genomic sequences that encode the novel splice form, nucleic acid variation (allelic information), tissue distribution of expression, and information about the closest art known protein/peptide/domain that has structural or sequence homology to the splice form of the present invention.

[0022] In addition to being previously unknown, the peptides that are provided in the present invention are selected based on their ability to be used for the development of commercially important products and services. Specifically, the present peptides are selected based on homology and/or structural relatedness to known secreted proteins of the nephronectin protein subfamily and the expression pattern observed. Experimental data as provided in FIG. 1 indicates expression in head/neck, nervous tumor, colon, breast, and placenta tissue. The art has clearly established the commercial importance of members of this family of proteins and proteins that have expression patterns similar to that of the present gene. Some of the more specific features of the peptides of the present invention, and the uses thereof, are described herein, particularly in the Background of the Invention and in the annotation provided in the Figures, and/or are known within the art for each of the known nephronectin family or subfamily of secreted proteins.

[0023] Specific Embodiments

[0024] Peptide Molecules

[0025] The present invention provides nucleic acid sequences that encode protein molecules that have been identified as being members of the secreted protein family of proteins and are related to the nephronectin protein subfamily (protein sequences are provided in FIG. 2, transcript/cDNA sequences are provided in FIG. 1 and genomic sequences are provided in FIG. 3). The peptide sequences provided in FIG. 2, as well as the obvious variants described herein, particularly allelic variants as identified herein and using the information in FIG. 3, will be referred herein as the secreted peptides of the present invention, secreted peptides, or peptides/proteins of the present invention.

[0026] The present invention provides isolated peptide and protein molecules that consist of, consist essentially of, or comprise the amino acid sequences of the secreted peptides disclosed in the FIG. 2, (encoded by the nucleic acid molecule shown in FIG. 1, transcript/cDNA or FIG. 3, genomic sequence), as well as all obvious variants of these peptides that are within the art to make and use. Some of these variants are described in detail below.

[0027] As used herein, a peptide is said to be “isolated” or “purified” when it is substantially free of cellular material or free of chemical precursors or other chemicals. The peptides of the present invention can be purified to homogeneity or other degrees of purity. The level of purification will be based on the intended use. The critical feature is that the preparation allows for the desired function of the peptide, even if in the presence of considerable amounts of other components (the features of an isolated nucleic acid molecule is discussed below).

[0028] In some uses, “substantially free of cellular material” includes preparations of the peptide having less than about 30% (by dry weight) other proteins (i.e., contaminating protein), less than about 20% other proteins, less than about 10% other proteins, or less than about 5% other proteins. When the peptide is recombinantly produced, it can also be substantially free of culture medium, i.e., culture medium represents less than about 20% of the volume of the protein preparation.

[0029] The language “substantially free of chemical precursors or other chemicals” includes preparations of the peptide in which it is separated from chemical precursors or other chemicals that are involved in its synthesis. In one embodiment, the language “substantially free of chemical precursors or other chemicals” includes preparations of the secreted peptide having less than about 30% (by dry weight) chemical precursors or other chemicals, less than about 20% chemical precursors or other chemicals, less than about 10% chemical precursors or other chemicals, or less than about 5% chemical precursors or other chemicals.

[0030] The isolated secreted peptide can be purified from cells that naturally express it, purified from cells that have been altered to express it (recombinant), or synthesized using known protein synthesis methods. Experimental data as provided in FIG. 1 indicates expression in head/neck, nervous tumor, colon, breast, and placenta tissue. For example, a nucleic acid molecule encoding the secreted peptide is cloned into an expression vector, the expression vector introduced into a host cell and the protein expressed in the host cell. The protein can then be isolated from the cells by an appropriate purification scheme using standard protein purification techniques. Many of these techniques are described in detail below.

[0031] Accordingly, the present invention provides proteins that consist of the amino acid sequences provided in FIG. 2 (SEQ ID NO:2), for example, proteins encoded by the transcript/cDNA nucleic acid sequences shown in FIG. 1 (SEQ ID NO:1) and the genomic sequences provided in FIG. 3 (SEQ ID NO:3). The amino acid sequence of such a protein is provided in FIG. 2. A protein consists of an amino acid sequence when the amino acid sequence is the final amino acid sequence of the protein.

[0032] The present invention further provides proteins that consist essentially of the amino acid sequences provided in FIG. 2 (SEQ ID NO:2), for example, proteins encoded by the transcript/cDNA nucleic acid sequences shown in FIG. 1 (SEQ ID NO:1) and the genomic sequences provided in FIG. 3 (SEQ ID NO:3). A protein consists essentially of an amino acid sequence when such an amino acid sequence is present with only a few additional amino acid residues, for example from about 1 to about 100 or so additional residues, typically from 1 to about 20 additional residues in the final protein.

[0033] The present invention further provides proteins that comprise the amino acid sequences provided in FIG. 2 (SEQ ID NO:2), for example, proteins encoded by the transcript/cDNA nucleic acid sequences shown in FIG. 1 (SEQ ID NO:1) and the genomic sequences provided in FIG. 3 (SEQ ID NO:3). A protein comprises an amino acid sequence when the amino acid sequence is at least part of the final amino acid sequence of the protein. In such a fashion, the protein can be only the peptide or have additional amino acid molecules, such as amino acid residues (contiguous encoded sequence) that are naturally associated with it or heterologous amino acid residues/peptide sequences. Such a protein can have a few additional amino acid residues or can comprise several hundred or more additional amino acids. The preferred classes of proteins that are comprised of the secreted peptides of the present invention are the naturally occurring mature proteins. A brief description of how various types of these proteins can be made/isolated is provided below.

[0034] The secreted peptides of the present invention can be attached to heterologous sequences to form chimeric or fusion proteins. Such chimeric and fusion proteins comprise a secreted peptide operatively linked to a heterologous protein having an amino acid sequence not substantially homologous to the secreted peptide. “Operatively linked” indicates that the secreted peptide and the heterologous protein are fused in-frame. The heterologous protein can be fused to the N-terminus or C-terminus of the secreted peptide.

[0035] In some uses, the fusion protein does not affect the activity of the secreted peptide per se.

[0036] For example, the fusion protein can include, but is not limited to, enzymatic fusion proteins, for example beta-galactosidase fusions, yeast two-hybrid GAL fusions, poly-His fusions, MYC-tagged, HI-tagged and Ig fusions. Such fusion proteins, particularly poly-His fusions, can facilitate the purification of recombinant secreted peptide. In certain host cells (e.g., mammalian host cells), expression and/or secretion of a protein can be increased by using a heterologous signal sequence.

[0037] A chimeric or fusion protein can be produced by standard recombinant DNA techniques. For example, DNA fragments coding for the different protein sequences are ligated together in-frame in accordance with conventional techniques. In another embodiment, the fusion gene can be synthesized by conventional techniques including automated DNA synthesizers. Alternatively, PCR amplification of gene fragments can be carried out using anchor primers which give rise to complementary overhangs between two consecutive gene fragments which can subsequently be annealed and re-amplified to generate a chimeric gene sequence (see Ausubel et al, Current Protocols in Molecular Biology, 1992). Moreover, many expression vectors are commercially available that already encode a fusion moiety (e.g., a GST protein). A secreted peptide-encoding nucleic acid can be cloned into such an expression vector such that the fusion moiety is linked in-frame to the secreted peptide.

[0038] As mentioned above, the present invention also provides and enables obvious variants of the amino acid sequence of the proteins of the present invention, such as naturally occurring mature forms of the peptide, allelic/sequence variants of the peptides, non-naturally occurring recombinantly derived variants of the peptides, and orthologs and paralogs of the peptides. Such variants can readily be generated using art-known techniques in the fields of recombinant nucleic acid technology and protein biochemistry. It is understood, however, that variants exclude any amino acid sequences disclosed prior to the invention.

[0039] Such variants can readily be identified/made using molecular techniques and the sequence information disclosed herein. Further, such variants can readily be distinguished from other peptides based on sequence and/or structural homology to the secreted peptides of the present invention. The degree of homology/identity present will be based primarily on whether the peptide is a functional variant or non-functional variant, the amount of divergence present in the paralog family and the evolutionary distance between the orthologs.

[0040] To determine the percent identity of two amino acid sequences or two nucleic acid sequences, the sequences are aligned for optimal comparison purposes (e.g., gaps can be introduced in one or both of a first and a second amino acid or nucleic acid sequence for optimal alignment and non-homologous sequences can be disregarded for comparison purposes). In a preferred embodiment, at least 30%, 40%, 50%, 60%, 70%, 80%, or 90% or more of the length of a reference sequence is aligned for comparison purposes. The amino acid residues or nucleotides at corresponding amino acid positions or nucleotide positions are then compared. When a position in the first sequence is occupied by the same amino acid residue or nucleotide as the corresponding position in the second sequence, then the molecules are identical at that position (as used herein amino acid or nucleic acid “identity” is equivalent to amino acid or nucleic acid “homology”). The percent identity between the two sequences is a function of the number of identical positions shared by the sequences, taking into account the number of gaps, and the length of each gap, which need to be introduced for optimal alignment of the two sequences.

[0041] The comparison of sequences and determination of percent identity and similarity between two sequences can be accomplished using a mathematical algorithm. (Computational Molecular Biology, Lesk, A. M., ed., Oxford University Press, New York, 1988; Biocomputing: Informatics and Genome Projects, Smith, D. W., ed., Academic Press, New York, 1993; Computer Analysis of Sequence Data, Part 1, Griffin, A. M., and Griffin, H. G., eds., Humana Press, New Jersey, 1994; Sequence Analysis in Molecular Biology, von Heinje, G., Academic Press, 1987; and Sequence Analysis Primer, Gribskov, M. and Devereux, J., eds., M Stockton Press, New York, 1991). In a preferred embodiment, the percent identity between two amino acid sequences is determined using the Needleman and Wunsch (J. Mol. Biol. (48):444-453 (1970)) algorithm which has been incorporated into the GAP program in the GCG software package (available at http://www.gcg.com), using either a Blossom 62 matrix or a PAM250 matrix, and a gap weight of 16, 14, 12, 10, 8, 6, or 4 and a length weight of 1, 2, 3, 4, 5, or 6. In yet another preferred embodiment, the percent identity between two nucleotide sequences is determined using the GAP program in the GCG software package (Devereux, J., et al., Nucleic Acids Res. 12(1):387 (1984)) (available at http://www.gcg.com), using a NWSgapdna.CMP matrix and a gap weight of 40, 50, 60, 70, or 80 and a length weight of 1, 2, 3, 4, 5, or 6. In another embodiment, the percent identity between two amino acid or nucleotide sequences is determined using the algorithm of E. Myers and W. Miller (CABIOS, 4:11-17 (1989)) which has been incorporated into the ALIGN program (version 2.0), using a PAM120 weight residue table, a gap length penalty of 12 and a gap penalty of 4.

[0042] The nucleic acid and protein sequences of the present invention can further be used as a “query sequence” to perform a search against sequence databases to, for example, identify other family members or related sequences. Such searches can be performed using the NBLAST and XBLAST programs (version 2.0) of Altschul, et al. (J. Mol. Biol. 215:403-10 (1990)). BLAST nucleotide searches can be performed with the NBLAST program, score=100, wordlength=12 to obtain nucleotide sequences homologous to the nucleic acid molecules of the invention. BLAST protein searches can be performed with the XBLAST program, score=50, wordlength=3 to obtain amino acid sequences homologous to the proteins of the invention. To obtain gapped alignments for comparison purposes, Gapped BLAST can be utilized as described in Altschul et al. (Nucleic Acids Res. 25(17):3389-3402 (1997)). When utilizing BLAST and gapped BLAST programs, the default parameters of the respective programs (e.g., XBLAST and NBLAST) can be used.

[0043] Full-length pre-processed forms, as well as mature processed forms, of proteins that comprise one of the peptides of the present invention can readily be identified as having complete sequence identity to one of the secreted peptides of the present invention as well as being encoded by the same genetic locus as the secreted peptide provided herein. As indicated by the data presented in FIG. 3, the map position was determined to be on human chromosome 4 by ePCR.

[0044] Allelic variants of a secreted peptide can readily be identified as being a human protein having a high degree (significant) of sequence homology/identity to at least a portion of the secreted peptide as well as being encoded by the same genetic locus as the secreted peptide provided herein. Genetic locus can readily be determined based on the genomic information provided in FIG. 3, such as the genomic sequence mapped to the reference human. As indicated by the data presented in FIG. 3, the map position was determined to be on human chromosome 4 by ePCR. As used herein, two proteins (or a region of the proteins) have significant homology when the amino acid sequences are typically at least about 70-80%, 80-90%, and more typically at least about 90-95% or more homologous. A significantly homologous amino acid sequence, according to the present invention, will be encoded by a nucleic acid sequence that will hybridize to a secreted peptide encoding nucleic acid molecule under stringent conditions as more fully described below.

[0045]FIG. 3 provides information on SNPs that have been found at 24 different nucleotide positions in the gene encoding the nephronectin protein of the present invention. Amongst the SNP variations was a non-synonymous coding SNP identified at nucleotide position 47,666 (protein position 281). The change in the amino acid sequence caused by this SNP is indicated in FIG. 3 and can readily be determined using the universal genetic code and the protein sequence provided in FIG. 2 as a reference.

[0046] Paralogs of a secreted peptide can readily be identified as having some degree of significant sequence homology/identity to at least a portion of the secreted peptide, as being encoded by a gene from humans, and as having similar activity or function. Two proteins will typically be considered paralogs when the amino acid sequences are typically at least about 60% or greater, and more typically at least about 70% or greater homology through a given region or domain. Such paralogs will be encoded by a nucleic acid sequence that will hybridize to a secreted peptide encoding nucleic acid molecule under moderate to stringent conditions as more fully described below.

[0047] Orthologs of a secreted peptide can readily be identified as having some degree of significant sequence homology/identity to at least a portion of the secreted peptide as well as being encoded by a gene from another organism. Preferred orthologs will be isolated from mammals, preferably primates, for the development of human therapeutic targets and agents. Such orthologs will be encoded by a nucleic acid sequence that will hybridize to a secreted peptide encoding nucleic acid molecule under moderate to stringent conditions, as more fully described below, depending on the degree of relatedness of the two organisms yielding the proteins.

[0048] Non-naturally occurring variants of the secreted peptides of the present invention can readily be generated using recombinant techniques. Such variants include, but are not limited to deletions, additions and substitutions in the amino acid sequence of the secreted peptide. For example, one class of substitutions are conserved amino acid substitution. Such substitutions are those that substitute a given amino acid in a secreted peptide by another amino acid of like characteristics. Typically seen as conservative substitutions are the replacements, one for another, among the aliphatic amino acids Ala, Val, Leu, and Ile; interchange of the hydroxyl residues Ser and Thr; exchange of the acidic residues Asp and Glu; substitution between the amide residues Asn and Gln; exchange of the basic residues Lys and Arg; and replacements among the aromatic residues Phe and Tyr. Guidance concerning which amino acid changes are likely to be phenotypically silent are found in Bowie et al., Science 247:1306-1310 (1990).

[0049] Variant secreted peptides can be fully functional or can lack function in one or more activities, e.g. ability to bind substrate, ability to phosphorylate substrate, ability to mediate signaling, etc. Fully functional variants typically contain only conservative variation or variation in non-critical residues or in non-critical regions. FIG. 2 provides the result of protein analysis and can be used to identify critical domains/regions. Functional variants can also contain substitution of similar amino acids that result in no change or an insignificant change in function. Alternatively, such substitutions may positively or negatively affect function to some degree.

[0050] Non-functional variants typically contain one or more non-conservative amino acid substitutions, deletions, insertions, inversions, or truncation or a substitution, insertion, inversion, or deletion in a critical residue or critical region.

[0051] Amino acids that are essential for function can be identified by methods known in the art, such as site-directed mutagenesis or alanine-scanning mutagenesis (Cunningham et al., Science 244:1081-1085 (1989)), particularly using the results provided in FIG. 2. The latter procedure introduces single alanine mutations at every residue in the molecule. The resulting mutant molecules are then tested for biological activity such as secreted protein activity or in assays such as an in vitro proliferative activity. Sites that are critical for binding partner/substrate binding can also be determined by structural analysis such as crystallization, nuclear magnetic resonance or photoaffinity labeling (Smith et al, J. Mol. Biol. 224:899-904 (1992); de Vos et al. Science 255:306-312 (1992)).

[0052] The present invention further provides fragments of the secreted peptides, in addition to proteins and peptides that comprise and consist of such fragments, particularly those comprising the residues identified in FIG. 2. The fragments to which the invention pertains, however, are not to be construed as encompassing fragments that may be disclosed publicly prior to the present invention.

[0053] As used herein, a fragment comprises at least 8, 10, 12, 14, 16, or more contiguous amino acid residues from a secreted peptide. Such fragments can be chosen based on the ability to retain one or more of the biological activities of the secreted peptide or could be chosen for the ability to perform a function, e.g. bind a substrate or act as an immunogen. Particularly important fragments are biologically active fragments, peptides that are, for example, about 8 or more amino acids in length. Such fragments will typically comprise a domain or motif of the secreted peptide, e.g., active site or a substrate-binding domain. Further, possible fragments include, but are not limited to, domain or motif containing fragments, soluble peptide fragments, and fragments containing immunogenic structures. Predicted domains and functional sites are readily identifiable by computer programs well known and readily available to those of skill in the art (e.g., PROSITE analysis). The results of one such analysis are provided in FIG. 2.

[0054] Polypeptides often contain amino acids other than the 20 amino acids commonly referred to as the 20 naturally occurring amino acids. Further, many amino acids, including the terminal amino acids, may be modified by natural processes, such as processing and other post-translational modifications, or by chemical modification techniques well known in the art. Common modifications that occur naturally in secreted peptides are described in basic texts, detailed monographs, and the research literature, and they are well known to those of skill in the art (some of these features are identified in FIG. 2).

[0055] Known modifications include, but are not limited to, acetylation, acylation, ADP-ribosylation, amidation, covalent attachment of flavin, covalent attachment of a heme moiety, covalent attachment of a nucleotide or nucleotide derivative, covalent attachment of a lipid or lipid derivative, covalent attachment of phosphotidylinositol, cross-linking, cyclization, disulfide bond formation, demethylation, formation of covalent crosslinks, formation of cystine, formation of pyroglutamate, formylation, gamma carboxylation, glycosylation, GPI anchor formation, hydroxylation, iodination, methylation, myristoylation, oxidation, proteolytic processing, phosphorylation, prenylation, racemization, selenoylation, sulfation, transfer-RNA mediated addition of amino acids to proteins such as arginylation, and ubiquitination.

[0056] Such modifications are well known to those of skill in the art and have been described in great detail in the scientific literature. Several particularly common modifications, glycosylation, lipid attachment, sulfation, gamma-carboxylation of glutamic acid residues, hydroxylation and ADP-ribosylation, for instance, are described in most basic texts, such as Proteins—Structure and Molecular Properties, 2nd Ed., T. E. Creighton, W. H. Freeman and Company, New York (1993). Many detailed reviews are available on this subject, such as by Wold, F., Posttranslational Covalent Modification of Proteins, B. C. Johnson, Ed., Academic Press, New York 1-12 (1983); Seifier et al. (Meth. Enzymol. 182: 626-646 (1990)) and Rattan et al (Ann. N.Y. Acad. Sci. 663:48-62 (1992)).

[0057] Accordingly, the secreted peptides of the present invention also encompass derivatives or analogs in which a substituted amino acid residue is not one encoded by the genetic code, in which a substituent group is included, in which the mature secreted peptide is fused with another compound, such as a compound to increase the half-life of the secreted peptide (for example, polyethylene glycol), or in which the additional amino acids are fused to the mature secreted peptide, such as a leader or secretory sequence or a sequence for purification of the mature secreted peptide or a pro-protein sequence.

[0058] Protein/Peptide Uses

[0059] The proteins of the present invention can be used in substantial and specific assays related to the functional information provided in the Figures; to raise antibodies or to elicit another immune response; as a reagent (including the labeled reagent) in assays designed to quantitatively determine levels of the protein (or its binding partner or ligand) in biological fluids; and as markers for tissues in which the corresponding protein is preferentially expressed (either constitutively or at a particular stage of tissue differentiation or development or in a disease state). Where the protein binds or potentially binds to another protein or ligand (such as, for example, in a secreted protein-effector protein interaction or secreted protein-ligand interaction), the protein can be used to identify the binding partner/ligand so as to develop a system to identify inhibitors of the binding interaction. Any or all of these uses are capable of being developed into reagent grade or kit format for commercialization as commercial products.

[0060] Methods for performing the uses listed above are well known to those skilled in the art. References disclosing such methods include “Molecular Cloning: A Laboratory Manual”, 2d ed., Cold Spring Harbor Laboratory Press, Sambrook, J., E. F. Fritsch and T. Maniatis eds., 1989, and “Methods in Enzymology: Guide to Molecular Cloning Techniques”, Academic Press, Berger, S. L. and A. R. Kimmel eds., 1987.

[0061] The potential uses of the peptides of the present invention are based primarily on the source of the protein as well as the class/action of the protein. For example, secreted proteins isolated from humans and their human/mammalian orthologs serve as targets for identifying agents for use in mammalian therapeutic applications, e.g. a human drug, particularly in modulating a biological or pathological response in a cell or tissue that expresses the secreted protein. Experimental data as provided in FIG. 1 indicates that secreted proteins of the present invention are expressed in head/neck, nervous tumor, colon, breast, and placenta tissue, as indicated by virtual northern blot analysis. A large percentage of pharmaceutical agents are being developed that modulate the activity of secreted proteins, particularly members of the nephronectin subfamily (see Background of the Invention). The structural and functional information provided in the Background and Figures provide specific and substantial uses for the molecules of the present invention, particularly in combination with the expression information provided in FIG. 1. Experimental data as provided in FIG. 1 indicates expression in head/neck, nervous tumor, colon, breast, and placenta tissue. Such uses can readily be determined using the information provided herein, that which is known in the art, and routine experimentation.

[0062] The proteins of the present invention (including variants and fragments that may have been disclosed prior to the present invention) are useful for biological assays related to secreted proteins that are related to members of the nephronectin subfamily. Such assays involve any of the known secreted protein functions or activities or properties useful for diagnosis and treatment of secreted protein-related conditions that are specific for the subfamily of secreted proteins that the one of the present invention belongs to, particularly in cells and tissues that express the secreted protein. Experimental data as provided in FIG. 1 indicates that secreted proteins of the present invention are expressed in head/neck, nervous tumor, colon, breast, and placenta tissue, as indicated by virtual northern blot analysis.

[0063] The proteins of the present invention are also useful in drug screening assays, in cell-based or cell-free systems. Cell-based systems can be native, i.e., cells that normally express the secreted protein, as a biopsy or expanded in cell culture. Experimental data as provided in FIG. 1 indicates expression in head/neck, nervous tumor, colon, breast, and placenta tissue. In an alternate embodiment, cell-based assays involve recombinant host cells expressing the secreted protein.

[0064] The polypeptides can be used to identify compounds that modulate secreted protein activity of the protein in its natural state or an altered form that causes a specific disease or pathology associated with the secreted protein. Both the secreted proteins of the present invention and appropriate variants and fragments can be used in high-throughput screens to assay candidate compounds for the ability to bind to the secreted protein. These compounds can be further screened against a functional secreted protein to determine the effect of the compound on the secreted protein activity. Further, these compounds can be tested in animal or invertebrate systems to determine activity/effectiveness. Compounds can be identified that activate (agonist) or inactivate (antagonist) the secreted protein to a desired degree.

[0065] Further, the proteins of the present invention can be used to screen a compound for the ability to stimulate or inhibit interaction between the secreted protein and a molecule that normally interacts with the secreted protein, e.g. a substrate or a component of the signal pathway that the secreted protein normally interacts (for example, another secreted protein). Such assays typically include the steps of combining the secreted protein with a candidate compound under conditions that allow the secreted protein, or fragment, to interact with the target molecule, and to detect the formation of a complex between the protein and the target or to detect the biochemical consequence of the interaction with the secreted protein and the target.

[0066] Candidate compounds include, for example, 1) peptides such as soluble peptides, including Ig-tailed fusion peptides and members of random peptide libraries (see, e.g., Lam et al., Nature 354:82-84 (1991); Houghten et al., Nature 354:84-86 (1991)) and combinatorial chemistry-derived molecular libraries made of D- and/or L-configuration amino acids; 2) phosphopeptides (e.g., members of random and partially degenerate, directed phosphopeptide libraries, see, e.g., Songyang et al., Cell 72:767-778 (1993)); 3) antibodies (e.g., polyclonal, monoclonal, humanized, anti-idiotypic, chimeric, and single chain antibodies as well as Fab, F(ab′)₂, Fab expression library fragments, and epitope-binding fragments of antibodies); and 4) small organic and inorganic molecules (e.g., molecules obtained from combinatorial and natural product libraries).

[0067] One candidate compound is a soluble fragment of the receptor that competes for substrate binding. Other candidate compounds include mutant secreted proteins or appropriate fragments containing mutations that affect secreted protein function and thus compete for substrate. Accordingly, a fragment that competes for substrate, for example with a higher affinity, or a fragment that binds substrate but does not allow release, is encompassed by the invention.

[0068] Any of the biological or biochemical functions mediated by the secreted protein can be used as an endpoint assay. These include all of the biochemical or biochemical/biological events described herein, in the references cited herein, incorporated by reference for these endpoint assay targets, and other functions known to those of ordinary skill in the art or that can be readily identified using the information provided in the Figures, particularly FIG. 2. Specifically, a biological function of a cell or tissues that expresses the secreted protein can be assayed. Experimental data as provided in FIG. 1 indicates that secreted proteins of the present invention are expressed in head/neck, nervous tumor, colon, breast, and placenta tissue, as indicated by virtual northern blot analysis.

[0069] Binding and/or activating compounds can also be screened by using chimeric secreted proteins in which the amino terminal extracellular domain, or parts thereof, the entire transmembrane domain or subregions, such as any of the seven transmembrane segments or any of the intracellular or extracellular loops and the carboxy terminal intracellular domain, or parts thereof, can be replaced by heterologous domains or subregions. For example, a substrate-binding region can be used that interacts with a different substrate then that which is recognized by the native secreted protein. Accordingly, a different set of signal transduction components is available as an end-point assay for activation. This allows for assays to be performed in other than the specific host cell from which the secreted protein is derived.

[0070] The proteins of the present invention are also useful in competition binding assays in methods designed to discover compounds that interact with the secreted protein (e.g. binding partners and/or ligands). Thus, a compound is exposed to a secreted protein polypeptide under conditions that allow the compound to bind or to otherwise interact with the polypeptide. Soluble secreted protein polypeptide is also added to the mixture. If the test compound interacts with the soluble secreted protein polypeptide, it decreases the amount of complex formed or activity from the secreted protein target. This type of assay is particularly useful in cases in which compounds are sought that interact with specific regions of the secreted protein. Thus, the soluble polypeptide that competes with the target secreted protein region is designed to contain peptide sequences corresponding to the region of interest.

[0071] To perform cell free drug screening assays, it is sometimes desirable to immobilize either the secreted protein, or fragment, or its target molecule to facilitate separation of complexes from uncomplexed forms of one or both of the proteins, as well as to accommodate automation of the assay.

[0072] Techniques for immobilizing proteins on matrices can be used in the drug screening assays. In one embodiment, a fusion protein can be provided which adds a domain that allows the protein to be bound to a matrix. For example, glutathione-S-transferase fusion proteins can be adsorbed onto glutathione sepharose beads (Sigma Chemical, St. Louis, Mo.) or glutathione derivatized microtitre plates, which are then combined with the cell lysates (e.g., ³⁵S-labeled) and the candidate compound, and the mixture incubated under conditions conducive to complex formation (e.g., at physiological conditions for salt and pH). Following incubation, the beads are washed to remove any unbound label, and the matrix immobilized and radiolabel determined directly, or in the supernatant after the complexes are dissociated. Alternatively, the complexes can be dissociated from the matrix, separated by SDS-PAGE, and the level of secreted protein-binding protein found in the bead fraction quantitated from the gel using standard electrophoretic techniques. For example, either the polypeptide or its target molecule can be immobilized utilizing conjugation of biotin and streptavidin using techniques well known in the art. Alternatively, antibodies reactive with the protein but which do not interfere with binding of the protein to its target molecule can be derivatized to the wells of the plate, and the protein trapped in the wells by antibody conjugation. Preparations of a secreted protein-binding protein and a candidate compound are incubated in the secreted protein-presenting wells and the amount of complex trapped in the well can be quantitated. Methods for detecting such complexes, in addition to those described above for the GST-immobilized complexes, include immunodetection of complexes using antibodies reactive with the secreted protein target molecule, or which are reactive with secreted protein and compete with the target molecule, as well as enzyme-linked assays which rely on detecting an enzymatic activity associated with the target molecule.

[0073] Agents that modulate one of the secreted proteins of the present invention can be identified using one or more of the above assays, alone or in combination. It is generally preferable to use a cell-based or cell free system first and then confirm activity in an animal or other model system. Such model systems are well known in the art and can readily be employed in this context.

[0074] Modulators of secreted protein activity identified according to these drug screening assays can be used to treat a subject with a disorder mediated by the secreted protein pathway, by treating cells or tissues that express the secreted protein. Experimental data as provided in FIG. 1 indicates expression in head/neck, nervous tumor, colon, breast, and placenta tissue. These methods of treatment include the steps of administering a modulator of secreted protein activity in a pharmaceutical composition to a subject in need of such treatment, the modulator being identified as described herein.

[0075] In yet another aspect of the invention, the secreted proteins can be used as “bait proteins” in a two-hybrid assay or three-hybrid assay (see, e.g., U.S. Pat. No. 5,283,317; Zervos et al. (1993) Cell 72:223-232; Madura et al. (1993) J. Biol. Chem. 268:12046-12054; Bartel et al. (1993) Biotechniques 14:920-924; Iwabuchi et al. (1993) Oncogene 8:1693-1696; and Brent WO94/10300), to identify other proteins, which bind to or interact with the secreted protein and are involved in secreted protein activity.

[0076] The two-hybrid system is based on the modular nature of most transcription factors, which consist of separable DNA-binding and activation domains. Briefly, the assay utilizes two different DNA constructs. In one construct, the gene that codes for a secreted protein is fused to a gene encoding the DNA binding domain of a known transcription factor (e.g., GAL-4). In the other construct, a DNA sequence, from a library of DNA sequences, that encodes an unidentified protein (“prey” or “sample”) is fused to a gene that codes for the activation domain of the known transcription factor. If the “bait” and the “prey” proteins are able to interact, in vivo, forming a secreted protein-dependent complex, the DNA-binding and activation domains of the transcription factor are brought into close proximity. This proximity allows transcription of a reporter gene (e.g., LacZ) which is operably linked to a transcriptional regulatory site responsive to the transcription factor. Expression of the reporter gene can be detected and cell colonies containing the functional transcription factor can be isolated and used to obtain the cloned gene which encodes the protein which interacts with the secreted protein.

[0077] This invention further pertains to novel agents identified by the above-described screening assays. Accordingly, it is within the scope of this invention to further use an agent identified as described herein in an appropriate animal model. For example, an agent identified as described herein (e.g., a secreted protein-modulating agent, an antisense secreted protein nucleic acid molecule, a secreted protein-specific antibody, or a secreted protein-binding partner) can be used in an animal or other model to determine the efficacy, toxicity, or side effects of treatment with such an agent. Alternatively, an agent identified as described herein can be used in an animal or other model to determine the mechanism of action of such an agent. Furthermore, this invention pertains to uses of novel agents identified by the above-described screening assays for treatments as described herein.

[0078] The secreted proteins of the present invention are also useful to provide a target for diagnosing a disease or predisposition to disease mediated by the peptide. Accordingly, the invention provides methods for detecting the presence, or levels of, the protein (or encoding mRNA) in a cell, tissue, or organism. Experimental data as provided in FIG. 1 indicates expression in head/neck, nervous tumor, colon, breast, and placenta tissue. The method involves contacting a biological sample with a compound capable of interacting with the secreted protein such that the interaction can be detected. Such an assay can be provided in a single detection format or a multi-detection format such as an antibody chip array.

[0079] One agent for detecting a protein in a sample is an antibody capable of selectively binding to protein. A biological sample includes tissues, cells and biological fluids isolated from a subject, as well as tissues, cells and fluids present within a subject.

[0080] The peptides of the present invention also provide targets for diagnosing active protein activity, disease, or predisposition to disease, in a patient having a variant peptide, particularly activities and conditions that are known for other members of the family of proteins to which the present one belongs. Thus, the peptide can be isolated from a biological sample and assayed for the presence of a genetic mutation that results in aberrant peptide. This includes amino acid substitution, deletion, insertion, rearrangement, (as the result of aberrant splicing events), and inappropriate post-translational modification. Analytic methods include altered electrophoretic mobility, altered tryptic peptide digest, altered secreted protein activity in cell-based or cell-free assay, alteration in substrate or antibody-binding pattern, altered isoelectric point, direct amino acid sequencing, and any other of the known assay techniques useful for detecting mutations in a protein. Such an assay can be provided in a single detection format or a multi-detection format such as an antibody chip array.

[0081] In vitro techniques for detection of peptide include enzyme linked immunosorbent assays (ELISAs), Western blots, immunoprecipitations and immunofluorescence using a detection reagent, such as an antibody or protein binding agent. Alternatively, the peptide can be detected in vivo in a subject by introducing into the subject a labeled anti-peptide antibody or other types of detection agent. For example, the antibody can be labeled with a radioactive marker whose presence and location in a subject can be detected by standard imaging techniques. Particularly useful are methods that detect the allelic variant of a peptide expressed in a subject and methods which detect fragments of a peptide in a sample.

[0082] The peptides are also useful in pharmacogenomic analysis. Pharmacogenomics deal with clinically significant hereditary variations in the response to drugs due to altered drug disposition and abnormal action in affected persons. See, e.g., Eichelbaum, M. (Clin. Exp. Pharmacol. Physiol. 23(10-11):983-985 (1996)), and Linder, M. W. (Clin. Chem. 43(2):254-266 (1997)). The clinical outcomes of these variations result in severe toxicity of therapeutic drugs in certain individuals or therapeutic failure of drugs in certain individuals as a result of individual variation in metabolism. Thus, the genotype of the individual can determine the way a therapeutic compound acts on the body or the way the body metabolizes the compound. Further, the activity of drug metabolizing enzymes effects both the intensity and duration of drug action. Thus, the pharmacogenomics of the individual permit the selection of effective compounds and effective dosages of such compounds for prophylactic or therapeutic treatment based on the individual's genotype. The discovery of genetic polymorphisms in some drug metabolizing enzymes has explained why some patients do not obtain the expected drug effects, show an exaggerated drug effect, or experience serious toxicity from standard drug dosages. Polymorphisms can be expressed in the phenotype of the extensive metabolizer and the phenotype of the poor metabolizer. Accordingly, genetic polymorphism may lead to allelic protein variants of the secreted protein in which one or more of the secreted protein functions in one population is different from those in another population. The peptides thus allow a target to ascertain a genetic predisposition that can affect treatment modality. Thus, in a ligand-based treatment, polymorphism may give rise to amino terminal extracellular domains and/or other substrate-binding regions that are more or less active in substrate binding, and secreted protein activation. Accordingly, substrate dosage would necessarily be modified to maximize the therapeutic effect within a given population containing a polymorphism. As an alternative to genotyping, specific polymorphic peptides could be identified.

[0083] The peptides are also useful for treating a disorder characterized by an absence of, inappropriate, or unwanted expression of the protein. Experimental data as provided in FIG. 1 indicates expression in head/neck, nervous tumor, colon, breast, and placenta tissue. Accordingly, methods for treatment include the use of the secreted protein or fragments.

[0084] Antibodies

[0085] The invention also provides antibodies that selectively bind to one of the peptides of the present invention, a protein comprising such a peptide, as well as variants and fragments thereof. As used herein, an antibody selectively binds a target peptide when it binds the target peptide and does not significantly bind to unrelated proteins. An antibody is still considered to selectively bind a peptide even if it also binds to other proteins that are not substantially homologous with the target peptide so long as such proteins share homology with a fragment or domain of the peptide target of the antibody. In this case, it would be understood that antibody binding to the peptide is still selective despite some degree of cross-reactivity.

[0086] As used herein, an antibody is defined in terms consistent with that recognized within the art: they are multi-subunit proteins produced by a mammalian organism in response to an antigen challenge. The antibodies of the present invention include polyclonal antibodies and monoclonal antibodies, as well as fragments of such antibodies, including, but not limited to, Fab or F(ab′)₂, and Fv fragments.

[0087] Many methods are known for generating and/or identifying antibodies to a given target peptide. Several such methods are described by Harlow, Antibodies, Cold Spring Harbor Press, (1989).

[0088] In general, to generate antibodies, an isolated peptide is used as an immunogen and is administered to a mammalian organism, such as a rat, rabbit or mouse. The full-length protein, an antigenic peptide fragment or a fusion protein can be used. Particularly important fragments are those covering functional domains, such as the domains identified in FIG. 2, and domain of sequence homology or divergence amongst the family, such as those that can readily be identified using protein alignment methods and as presented in the Figures.

[0089] Antibodies are preferably prepared from regions or discrete fragments of the secreted proteins. Antibodies can be prepared from any region of the peptide as described herein. However, preferred regions will include those involved in function/activity and/or secreted protein/binding partner interaction. FIG. 2 can be used to identify particularly important regions while sequence alignment can be used to identify conserved and unique sequence fragments.

[0090] An antigenic fragment will typically comprise at least 8 contiguous amino acid residues. The antigenic peptide can comprise, however, at least 10, 12, 14, 16 or more amino acid residues. Such fragments can be selected on a physical property, such as fragments correspond to regions that are located on the surface of the protein, e.g., hydrophilic regions or can be selected based on sequence uniqueness (see FIG. 2).

[0091] Detection on an antibody of the present invention can be facilitated by coupling (i.e., physically linking) the antibody to a detectable substance. Examples of detectable substances include various enzymes, prosthetic groups, fluorescent materials, luminescent materials, bioluminescent materials, and radioactive materials. Examples of suitable enzymes include horseradish peroxidase, alkaline phosphatase, β-galactosidase, or acetylcholinesterase; examples of suitable prosthetic group complexes include streptavidin/biotin and avidin/biotin; examples of suitable fluorescent materials include umbelliferone, fluorescein, fluorescein isothiocyanate, rhodamine, dichlorotriazinylamine fluorescein, dansyl chloride or phycoerythrin; an example of a luminescent material includes luminol; examples of bioluminescent materials include luciferase, luciferin, and aequorin, and examples of suitable radioactive material include ¹²⁵I, ¹³¹I, ³⁵S or ³H.

[0092] Antibody Uses

[0093] The antibodies can be used to isolate one of the proteins of the present invention by standard techniques, such as affinity chromatography or immunoprecipitation. The antibodies can facilitate the purification of the natural protein from cells and recombinantly produced protein expressed in host cells. In addition, such antibodies are useful to detect the presence of one of the proteins of the present invention in cells or tissues to determine the pattern of expression of the protein among various tissues in an organism and over the course of normal development. Experimental data as provided in FIG. 1 indicates that secreted proteins of the present invention are expressed in head/neck, nervous tumor, colon, breast, and placenta tissue, as indicated by virtual northern blot analysis. Further, such antibodies can be used to detect protein in situ, in vitro, or in a cell lysate or supernatant in order to evaluate the abundance and pattern of expression. Also, such antibodies can be used to assess abnormal tissue distribution or abnormal expression during development or progression of a biological condition. Antibody detection of circulating fragments of the full length protein can be used to identify turnover.

[0094] Further, the antibodies can be used to assess expression in disease states such as in active stages of the disease or in an individual with a predisposition toward disease related to the protein's function. When a disorder is caused by an inappropriate tissue distribution, developmental expression, level of expression of the protein, or expressed/processed form, the antibody can be prepared against the normal protein. Experimental data as provided in FIG. 1 indicates expression in head/neck, nervous tumor, colon, breast, and placenta tissue. If a disorder is characterized by a specific mutation in the protein, antibodies specific for this mutant protein can be used to assay for the presence of the specific mutant protein.

[0095] The antibodies can also be used to assess normal and aberrant subcellular localization of cells in the various tissues in an organism. Experimental data as provided in FIG. 1 indicates expression in head/neck, nervous tumor, colon, breast, and placenta tissue. The diagnostic uses can be applied, not only in genetic testing, but also in monitoring a treatment modality. Accordingly, where treatment is ultimately aimed at correcting expression level or the presence of aberrant sequence and aberrant tissue distribution or developmental expression, antibodies directed against the protein or relevant fragments can be used to monitor therapeutic efficacy.

[0096] Additionally, antibodies are useful in pharmacogenomic analysis. Thus, antibodies prepared against polymorphic proteins can be used to identify individuals that require modified treatment modalities. The antibodies are also useful as diagnostic tools as an immunological marker for aberrant protein analyzed by electrophoretic mobility, isoelectric point, tryptic peptide digest, and other physical assays known to those in the art.

[0097] The antibodies are also useful for tissue typing. Experimental data as provided in FIG. 1 indicates expression in head/neck, nervous tumor, colon, breast, and placenta tissue. Thus, where a specific protein has been correlated with expression in a specific tissue, antibodies that are specific for this protein can be used to identify a tissue type.

[0098] The antibodies are also useful for inhibiting protein function, for example, blocking the binding of the secreted peptide to a binding partner such as a substrate. These uses can also be applied in a therapeutic context in which treatment involves inhibiting the protein's function. An antibody can be used, for example, to block binding, thus modulating (agonizing or antagonizing) the peptides activity. Antibodies can be prepared against specific fragments containing sites required for function or against intact protein that is associated with a cell or cell membrane. See FIG. 2 for structural information relating to the proteins of the present invention.

[0099] The invention also encompasses kits for using antibodies to detect the presence of a protein in a biological sample. The kit can comprise antibodies such as a labeled or labelable antibody and a compound or agent for detecting protein in a biological sample; means for determining the amount of protein in the sample; means for comparing the amount of protein in the sample with a standard; and instructions for use. Such a kit can be supplied to detect a single protein or epitope or can be configured to detect one of a multitude of epitopes, such as in an antibody detection array. Arrays are described in detail below for nuleic acid arrays and similar methods have been developed for antibody arrays.

[0100] Nucleic Acid Molecules

[0101] The present invention further provides isolated nucleic acid molecules that encode a secreted peptide or protein of the present invention (cDNA, transcript and genomic sequence). Such nucleic acid molecules will consist of, consist essentially of, or comprise a nucleotide sequence that encodes one of the secreted peptides of the present invention, an allelic variant thereof, or an ortholog or paralog thereof.

[0102] As used herein, an “isolated” nucleic acid molecule is one that is separated from other nucleic acid present in the natural source of the nucleic acid. Preferably, an “isolated” nucleic acid is free of sequences which naturally flank the nucleic acid (i.e., sequences located at the 5′ and 3′ ends of the nucleic acid) in the genomic DNA of the organism from which the nucleic acid is derived. However, there can be some flanking nucleotide sequences, for example up to about 5 KB, 4 KB, 3 KB, 2 KB, or 1 KB or less, particularly contiguous peptide encoding sequences and peptide encoding sequences within the same gene but separated by introns in the genomic sequence. The important point is that the nucleic acid is isolated from remote and unimportant flanking sequences such that it can be subjected to the specific manipulations described herein such as recombinant expression, preparation of probes and primers, and other uses specific to the nucleic acid sequences.

[0103] Moreover, an “isolated” nucleic acid molecule, such as a transcript/cDNA molecule, can be substantially free of other cellular material, or culture medium when produced by recombinant techniques, or chemical precursors or other chemicals when chemically synthesized. However, the nucleic acid molecule can be fused to other coding or regulatory sequences and still be considered isolated.

[0104] For example, recombinant DNA molecules contained in a vector are considered isolated. Further examples of isolated DNA molecules include recombinant DNA molecules maintained in heterologous host cells or purified (partially or substantially) DNA molecules in solution. Isolated RNA molecules include in vivo or in vitro RNA transcripts of the isolated DNA molecules of the present invention. Isolated nucleic acid molecules according to the present invention further include such molecules produced synthetically.

[0105] Accordingly, the present invention provides nucleic acid molecules that consist of the nucleotide sequence shown in FIG. 1 or 3 (SEQ ID NO:1, transcript sequence and SEQ ID NO:3, genomic sequence), or any nucleic acid molecule that encodes the protein provided in FIG. 2, SEQ ID NO:2. A nucleic acid molecule consists of a nucleotide sequence when the nucleotide sequence is the complete nucleotide sequence of the nucleic acid molecule.

[0106] The present invention further provides nucleic acid molecules that consist essentially of the nucleotide sequence shown in FIG. 1 or 3 (SEQ ID NO:1, transcript sequence and SEQ ID NO:3, genomic sequence), or any nucleic acid molecule that encodes the protein provided in FIG. 2, SEQ ID NO:2. A nucleic acid molecule consists essentially of a nucleotide sequence when such a nucleotide sequence is present with only a few additional nucleic acid residues in the final nucleic acid molecule.

[0107] The present invention further provides nucleic acid molecules that comprise the nucleotide sequences shown in FIG. 1 or 3 (SEQ ID NO:1, transcript sequence and SEQ ID NO:3, genomic sequence), or any nucleic acid molecule that encodes the protein provided in FIG. 2, SEQ ID NO:2. A nucleic acid molecule comprises a nucleotide sequence when the nucleotide sequence is at least part of the final nucleotide sequence of the nucleic acid molecule. In such a fashion, the nucleic acid molecule can be only the nucleotide sequence or have additional nucleic acid residues, such as nucleic acid residues that are naturally associated with it or heterologous nucleotide sequences. Such a nucleic acid molecule can have a few additional nucleotides or can comprises several hundred or more additional nucleotides. A brief description of how various types of these nucleic acid molecules can be readily made/isolated is provided below.

[0108] In FIGS. 1 and 3, both coding and non-coding sequences are provided. Because of the source of the present invention, humans genomic sequence (FIG. 3) and cDNA/transcript sequences (FIG. 1), the nucleic acid molecules in the Figures will contain genomic intronic sequences, 5′ and 3′ non-coding sequences, gene regulatory regions and non-coding intergenic sequences. In general such sequence features are either noted in FIGS. 1 and 3 or can readily be identified using computational tools known in the art. As discussed below, some of the non-coding regions, particularly gene regulatory elements such as promoters, are useful for a variety of purposes, e.g. control of heterologous gene expression, target for identifying gene activity modulating compounds, and are particularly claimed as fragments of the genomic sequence provided herein.

[0109] The isolated nucleic acid molecules can encode the mature protein plus additional amino or carboxyl-terminal amino acids, or amino acids interior to the mature peptide (when the mature form has more than one peptide chain, for instance). Such sequences may play a role in processing of a protein from precursor to a mature form, facilitate protein trafficking, prolong or shorten protein half-life or facilitate manipulation of a protein for assay or production, among other things. As generally is the case in situ, the additional amino acids may be processed away from the mature protein by cellular enzymes.

[0110] As mentioned above, the isolated nucleic acid molecules include, but are not limited to, the sequence encoding the secreted peptide alone, the sequence encoding the mature peptide and additional coding sequences, such as a leader or secretory sequence (e.g., a pre-pro or pro-protein sequence), the sequence encoding the mature peptide, with or without the additional coding sequences, plus additional non-coding sequences, for example introns and non-coding 5′ and 3′ sequences such as transcribed but non-translated sequences that play a role in transcription, mRNA processing (including splicing and polyadenylation signals), ribosome binding and stability of mRNA. In addition, the nucleic acid molecule may be fused to a marker sequence encoding, for example, a peptide that facilitates purification.

[0111] Isolated nucleic acid molecules can be in the form of RNA, such as mRNA, or in the form DNA, including cDNA and genomic DNA obtained by cloning or produced by chemical synthetic techniques or by a combination thereof. The nucleic acid, especially DNA, can be double-stranded or single-stranded. Single-stranded nucleic acid can be the coding strand (sense strand) or the non-coding strand (anti-sense strand).

[0112] The invention further provides nucleic acid molecules that encode fragments of the peptides of the present invention as well as nucleic acid molecules that encode obvious variants of the secreted proteins of the present invention that are described above. Such nucleic acid molecules may be naturally occurring, such as allelic variants (same locus), paralogs (different locus), and orthologs (different organism), or may be constructed by recombinant DNA methods or by chemical synthesis. Such non-naturally occurring variants may be made by mutagenesis techniques, including those applied to nucleic acid molecules, cells, or organisms. Accordingly, as discussed above, the variants can contain nucleotide substitutions, deletions, inversions and insertions. Variation can occur in either or both the coding and non-coding regions. The variations can produce both conservative and non-conservative amino acid substitutions.

[0113] The present invention further provides non-coding fragments of the nucleic acid molecules provided in FIGS. 1 and 3. Preferred non-coding fragments include, but are not limited to, promoter sequences, enhancer sequences, gene modulating sequences and gene termination sequences. Such fragments are useful in controlling heterologous gene expression and in developing screens to identify gene-modulating agents. A promoter can readily be identified as being 5′ to the ATG start site in the genomic sequence provided in FIG. 3.

[0114] A fragment comprises a contiguous nucleotide sequence greater than 12 or more nucleotides. Further, a fragment could at least 30, 40, 50, 100, 250 or 500 nucleotides in length. The length of the fragment will be based on its intended use. For example, the fragment can encode epitope bearing regions of the peptide, or can be useful as DNA probes and primers. Such fragments can be isolated using the known nucleotide sequence to synthesize an oligonucleotide probe. A labeled probe can then be used to screen a cDNA library, genomic DNA library, or mRNA to isolate nucleic acid corresponding to the coding region. Further, primers can be used in PCR reactions to clone specific regions of gene.

[0115] A probe/primer typically comprises substantially a purified oligonucleotide or oligonucleotide pair. The oligonucleotide typically comprises a region of nucleotide sequence that hybridizes under stringent conditions to at least about 12, 20, 25, 40, 50 or more consecutive nucleotides.

[0116] Orthologs, homologs, and allelic variants can be identified using methods well known in the art. As described in the Peptide Section, these variants comprise a nucleotide sequence encoding a peptide that is typically 60-70%, 70-80%, 80-90%, and more typically at least about 90-95% or more homologous to the nucleotide sequence shown in the Figure sheets or a fragment of this sequence. Such nucleic acid molecules can readily be identified as being able to hybridize under moderate to stringent conditions, to the nucleotide sequence shown in the Figure sheets or a fragment of the sequence. Allelic variants can readily be determined by genetic locus of the encoding gene. As indicated by the data presented in FIG. 3, the map position was determined to be on human chromosome 4 by ePCR.

[0117]FIG. 3 provides information on SNPs that have been found at 24 different nucleotide positions in the gene encoding the nephronectin protein of the present invention. Amongst the SNP variations was a non-synonymous coding SNP identified at nucleotide position 47,666 (protein position 281). The change in the amino acid sequence caused by this SNP is indicated in FIG. 3 and can readily be determined using the universal genetic code and the protein sequence provided in FIG. 2 as a reference.

[0118] As used herein, the term “hybridizes under stringent conditions” is intended to describe conditions for hybridization and washing under which nucleotide sequences encoding a peptide at least 60-70% homologous to each other typically remain hybridized to each other. The conditions can be such that sequences at least about 60%, at least about 70%, or at least about 80% or more homologous to each other typically remain hybridized to each other. Such stringent conditions are known to those skilled in the art and can be found in Current Protocols in Molecular Biology, John Wiley & Sons, N.Y. (1989), 6.3.1-6.3.6. One example of stringent hybridization conditions are hybridization in 6×sodium chloride/sodium citrate (SSC) at about 45C, followed by one or more washes in 0.2×SSC, 0.1% SDS at 50-65C. Examples of moderate to low stringency hybridization conditions are well known in the art.

[0119] Nucleic Acid Molecule Uses

[0120] The nucleic acid molecules of the present invention are useful for probes, primers, chemical intermediates, and in biological assays. The nucleic acid molecules are useful as a hybridization probe for messenger RNA, transcript/cDNA and genomic DNA to isolate full-length cDNA and genomic clones encoding the peptide described in FIG. 2 and to isolate cDNA and genomic clones that correspond to variants (alleles, orthologs, etc.) producing the same or related peptides shown in FIG. 2. As illustrated in FIG. 3, SNPs were identified at 24 different nucleotide positions in the nephronectin gene of the present invention.

[0121] The probe can correspond to any sequence along the entire length of the nucleic acid molecules provided in the Figures. Accordingly, it could be derived from 5′ noncoding regions, the coding region, and 3′ noncoding regions. However, as discussed, fragments are not to be construed as encompassing fragments disclosed prior to the present invention.

[0122] The nucleic acid molecules are also useful as primers for PCR to amplify any given region of a nucleic acid molecule and are useful to synthesize antisense molecules of desired length and sequence.

[0123] The nucleic acid molecules are also useful for constructing recombinant vectors. Such vectors include expression vectors that express a portion of, or all of, the peptide sequences. Vectors also include insertion vectors, used to integrate into another nucleic acid molecule sequence, such as into the cellular genome, to alter in situ expression of a gene and/or gene product. For example, an endogenous coding sequence can be replaced via homologous recombination with all or part of the coding region containing one or more specifically introduced mutations.

[0124] The nucleic acid molecules are also useful for expressing antigenic portions of the proteins.

[0125] The nucleic acid molecules are also useful as probes for determining the chromosomal positions of the nucleic acid molecules by means of in situ hybridization methods. As indicated by the data presented in FIG. 3, the map position was determined to be on human chromosome 4 by ePCR.

[0126] The nucleic acid molecules are also useful in making vectors containing the gene regulatory regions of the nucleic acid molecules of the present invention.

[0127] The nucleic acid molecules are also useful for designing ribozymes corresponding to all, or a part, of the mRNA produced from the nucleic acid molecules described herein.

[0128] The nucleic acid molecules are also useful for making vectors that express part, or all, of the peptides.

[0129] The nucleic acid molecules are also useful for constructing host cells expressing a part, or all, of the nucleic acid molecules and peptides.

[0130] The nucleic acid molecules are also useful for constructing transgenic animals expressing all, or a part, of the nucleic acid molecules and peptides.

[0131] The nucleic acid molecules are also useful as hybridization probes for determining the presence, level, form and distribution of nucleic acid expression. Experimental data as provided in FIG. 1 indicates that secreted proteins of the present invention are expressed in head/neck, nervous tumor, colon, breast, and placenta tissue, as indicated by virtual northern blot analysis. Accordingly, the probes can be used to detect the presence of, or to determine levels of, a specific nucleic acid molecule in cells, tissues, and in organisms. The nucleic acid whose level is determined can be DNA or RNA. Accordingly, probes corresponding to the peptides described herein can be used to assess expression and/or gene copy number in a given cell, tissue, or organism. These uses are relevant for diagnosis of disorders involving an increase or decrease in secreted protein expression relative to normal results.

[0132] In vitro techniques for detection of mRNA include Northern hybridizations and in situ hybridizations. In vitro techniques for detecting DNA include Southern hybridizations and in situ hybridization.

[0133] Probes can be used as a part of a diagnostic test kit for identifying cells or tissues that express a secreted protein, such as by measuring a level of a secreted protein-encoding nucleic acid in a sample of cells from a subject e.g., mRNA or genomic DNA, or determining if a secreted protein gene has been mutated. Experimental data as provided in FIG. 1 indicates that secreted proteins of the present invention are expressed in head/neck, nervous tumor, colon, breast, and placenta tissue, as indicated by virtual northern blot analysis.

[0134] Nucleic acid expression assays are useful for drug screening to identify compounds that modulate secreted protein nucleic acid expression.

[0135] The invention thus provides a method for identifying a compound that can be used to treat a disorder associated with nucleic acid expression of the secreted protein gene, particularly biological and pathological processes that are mediated by the secreted protein in cells and tissues that express it. Experimental data as provided in FIG. 1 indicates expression in head/neck, nervous tumor, colon, breast, and placenta tissue. The method typically includes assaying the ability of the compound to modulate the expression of the secreted protein nucleic acid and thus identifying a compound that can be used to treat a disorder characterized by undesired secreted protein nucleic acid expression. The assays can be performed in cell-based and cell-free systems. Cell-based assays include cells naturally expressing the secreted protein nucleic acid or recombinant cells genetically engineered to express specific nucleic acid sequences.

[0136] Thus, modulators of secreted protein gene expression can be identified in a method wherein a cell is contacted with a candidate compound and the expression of mRNA determined. The level of expression of secreted protein mRNA in the presence of the candidate compound is compared to the level of expression of secreted protein mRNA in the absence of the candidate compound. The candidate compound can then be identified as a modulator of nucleic acid expression based on this comparison and be used, for example to treat a disorder characterized by aberrant nucleic acid expression. When expression of mRNA is statistically significantly greater in the presence of the candidate compound than in its absence, the candidate compound is identified as a stimulator of nucleic acid expression. When nucleic acid expression is statistically significantly less in the presence of the candidate compound than in its absence, the candidate compound is identified as an inhibitor of nucleic acid expression.

[0137] The invention further provides methods of treatment, with the nucleic acid as a target, using a compound identified through drug screening as a gene modulator to modulate secreted protein nucleic acid expression in cells and tissues that express the secreted protein. Experimental data as provided in FIG. 1 indicates that secreted proteins of the present invention are expressed in head/neck, nervous tumor, colon, breast, and placenta tissue, as indicated by virtual northern blot analysis. Modulation includes both up-regulation (i.e. activation or agonization) or down-regulation (suppression or antagonization) or nucleic acid expression.

[0138] Alternatively, a modulator for secreted protein nucleic acid expression can be a small molecule or drug identified using the screening assays described herein as long as the drug or small molecule inhibits the secreted protein nucleic acid expression in the cells and tissues that express the protein. Experimental data as provided in FIG. 1 indicates expression in head/neck, nervous tumor, colon, breast, and placenta tissue.

[0139] The nucleic acid molecules are also useful for monitoring the effectiveness of modulating compounds on the expression or activity of the secreted protein gene in clinical trials or in a treatment regimen. Thus, the gene expression pattern can serve as a barometer for the continuing effectiveness of treatment with the compound, particularly with compounds to which a patient can develop resistance. The gene expression pattern can also serve as a marker indicative of a physiological response of the affected cells to the compound. Accordingly, such monitoring would allow either increased administration of the compound or the administration of alternative compounds to which the patient has not become resistant. Similarly, if the level of nucleic acid expression falls below a desirable level, administration of the compound could be commensurately decreased.

[0140] The nucleic acid molecules are also useful in diagnostic assays for qualitative changes in secreted protein nucleic acid expression, and particularly in qualitative changes that lead to pathology. The nucleic acid molecules can be used to detect mutations in secreted protein genes and gene expression products such as mRNA. The nucleic acid molecules can be used as hybridization probes to detect naturally occurring genetic mutations in the secreted protein gene and thereby to determine whether a subject with the mutation is at risk for a disorder caused by the mutation. Mutations include deletion, addition, or substitution of one or more nucleotides in the gene, chromosomal rearrangement, such as inversion or transposition, modification of genomic DNA, such as aberrant methylation patterns or changes in gene copy number, such as amplification. Detection of a mutated form of the secreted protein gene associated with a dysfunction provides a diagnostic tool for an active disease or susceptibility to disease when the disease results from overexpression, underexpression, or altered expression of a secreted protein.

[0141] Individuals carrying mutations in the secreted protein gene can be detected at the nucleic acid level by a variety of techniques. FIG. 3 provides information on SNPs that have been found at 24 different nucleotide positions in the gene encoding the nephronectin protein of the present invention. Amongst the SNP variations was a non-synonymous coding SNP identified at nucleotide position 47,666 (protein position 281). The change in the amino acid sequence caused by this SNP is indicated in FIG. 3 and can readily be determined using the universal genetic code and the protein sequence provided in FIG. 2 as a reference. As indicated by the data presented in FIG. 3, the map position was determined to be on human chromosome 4 by ePCR. Genomic DNA can be analyzed directly or can be amplified by using PCR prior to analysis. RNA or cDNA can be used in the same way. In some uses, detection of the mutation involves the use of a probe/primer in a polymerase chain reaction (PCR) (see, e.g. U.S. Pat. Nos. 4,683,195 and 4,683,202), such as anchor PCR or RACE PCR, or, alternatively, in a ligation chain reaction (LCR) (see, e.g., Landegran et al., Science 241:1077-1080 (1988); and Nakazawa et al., PNAS 91:360-364 (1994)), the latter of which can be particularly useful for detecting point mutations in the gene (see Abravaya et al., Nucleic Acids Res. 23:675-682 (1995)). This method can include the steps of collecting a sample of cells from a patient, isolating nucleic acid (e.g., genomic, mRNA or both) from the cells of the sample, contacting the nucleic acid sample with one or more primers which specifically hybridize to a gene under conditions such that hybridization and amplification of the gene (if present) occurs, and detecting the presence or absence of an amplification product, or detecting the size of the amplification product and comparing the length to a control sample. Deletions and insertions can be detected by a change in size of the amplified product compared to the normal genotype. Point mutations can be identified by hybridizing amplified DNA to normal RNA or antisense DNA sequences.

[0142] Alternatively, mutations in a secreted protein gene can be directly identified, for example, by alterations in restriction enzyme digestion patterns determined by gel electrophoresis.

[0143] Further, sequence-specific ribozymes (U.S. Pat. No. 5,498,531) can be used to score for the presence of specific mutations by development or loss of a ribozyme cleavage site. Perfectly matched sequences can be distinguished from mismatched sequences by nuclease cleavage digestion assays or by differences in melting temperature.

[0144] Sequence changes at specific locations can also be assessed by nuclease protection assays such as RNase and S1 protection or the chemical cleavage method. Furthermore, sequence differences between a mutant secreted protein gene and a wild-type gene can be determined by direct DNA sequencing. A variety of automated sequencing procedures can be utilized when performing the diagnostic assays (Naeve, C. W., (1995) Biotechniques 19:448), including sequencing by mass spectrometry (see, e.g., PCT International Publication No. WO 94/16101; Cohen et al., Adv. Chromatogr. 36:127-162 (1996); and Griffin et al., Appl. Biochem. Biotechnol. 38:147-159 (1993)).

[0145] Other methods for detecting mutations in the gene include methods in which protection from cleavage agents is used to detect mismatched bases in RNA/RNA or RNA/DNA duplexes (Myers et al., Science 230:1242 (1985)); Cotton et al., PNAS 85:4397 (1988); Saleeba et al., Meth. Enzymol. 217:286-295 (1992)), electrophoretic mobility of mutant and wild type nucleic acid is compared (Orita et al., PNAS 86:2766 (1989); Cotton et al., Mutat. Res. 285:125-144 (1993); and Hayashi et al., Genet. Anal. Tech. Appl. 9:73-79 (1992)), and movement of mutant or wild-type fragments in polyacrylamide gels containing a gradient of denaturant is assayed using denaturing gradient gel electrophoresis (Myers et al., Nature 313:495 (1985)). Examples of other techniques for detecting point mutations include selective oligonucleotide hybridization, selective amplification, and selective primer extension.

[0146] The nucleic acid molecules are also useful for testing an individual for a genotype that while not necessarily causing the disease, nevertheless affects the treatment modality. Thus, the nucleic acid molecules can be used to study the relationship between an individual's genotype and the individual's response to a compound used for treatment (pharmacogenomic relationship). Accordingly, the nucleic acid molecules described herein can be used to assess the mutation content of the secreted protein gene in an individual in order to select an appropriate compound or dosage regimen for treatment. FIG. 3 provides information on SNPs that have been found at 24 different nucleotide positions in the gene encoding the nephronectin protein of the present invention. Amongst the SNP variations was a non-synonymous coding SNP identified at nucleotide position 47,666 (protein position 281). The change in the amino acid sequence caused by this SNP is indicated in FIG. 3 and can readily be determined using the universal genetic code and the protein sequence provided in FIG. 2 as a reference.

[0147] Thus nucleic acid molecules displaying genetic variations that affect treatment provide a diagnostic target that can be used to tailor treatment in an individual. Accordingly, the production of recombinant cells and animals containing these polymorphisms allow effective clinical design of treatment compounds and dosage regimens.

[0148] The nucleic acid molecules are thus useful as antisense constructs to control secreted protein gene expression in cells, tissues, and organisms. A DNA antisense nucleic acid molecule is designed to be complementary to a region of the gene involved in transcription, preventing transcription and hence production of secreted protein. An antisense RNA or DNA nucleic acid molecule would hybridize to the mRNA and thus block translation of mRNA into secreted protein.

[0149] Alternatively, a class of antisense molecules can be used to inactivate mRNA in order to decrease expression of secreted protein nucleic acid. Accordingly, these molecules can treat a disorder characterized by abnormal or undesired secreted protein nucleic acid expression. This technique involves cleavage by means of ribozymes containing nucleotide sequences complementary to one or more regions in the mRNA that attenuate the ability of the mRNA to be translated. Possible regions include coding regions and particularly coding regions corresponding to the catalytic and other functional activities of the secreted protein, such as substrate binding.

[0150] The nucleic acid molecules also provide vectors for gene therapy in patients containing cells that are aberrant in secreted protein gene expression. Thus, recombinant cells, which include the patient's cells that have been engineered ex vivo and returned to the patient, are introduced into an individual where the cells produce the desired secreted protein to treat the individual.

[0151] The invention also encompasses kits for detecting the presence of a secreted protein nucleic acid in a biological sample. Experimental data as provided in FIG. 1 indicates that secreted proteins of the present invention are expressed in head/neck, nervous tumor, colon, breast, and placenta tissue, as indicated by virtual northern blot analysis. For example, the kit can comprise reagents such as a labeled or labelable nucleic acid or agent capable of detecting secreted protein nucleic acid in a biological sample; means for determining the amount of secreted protein nucleic acid in the sample; and means for comparing the amount of secreted protein nucleic acid in the sample with a standard. The compound or agent can be packaged in a suitable container. The kit can further comprise instructions for using the kit to detect secreted protein mRNA or DNA.

[0152] Nucleic Acid Arrays

[0153] The present invention further provides nucleic acid detection kits, such as arrays or microarrays of nucleic acid molecules that are based on the sequence information provided in FIGS. 1 and 3 (SEQ ID NOS:1 and 3). As used herein “Arrays” or “Microarrays” refers to an array of distinct polynucleotides or oligonucleotides synthesized on a substrate, such as paper, nylon or other type of membrane, filter, chip, glass slide, or any other suitable solid support. In one embodiment, the microarray is prepared and used according to the methods described in U.S. Pat. No. 5,837,832, Chee et al., PCT application WO95/11995 (Chee et al.), Lockhart, D. J. et al. (1996; Nat. Biotech. 14: 1675-1680) and Schena, M. et al. (1996; Proc. Natl. Acad. Sci. 93: 10614-10619), all of which are incorporated herein in their entirety by reference. In other embodiments, such arrays are produced by the methods described by Brown et al., U.S. Pat. No. 5,807,522.

[0154] The microarray or detection kit is preferably composed of a large number of unique, single-stranded nucleic acid sequences, usually either synthetic antisense oligonucleotides or fragments of cDNAs, fixed to a solid support. The oligonucleotides are preferably about 6-60 nucleotides in length, more preferably 15-30 nucleotides in length, and most preferably about 20-25 nucleotides in length. For a certain type of microarray or detection kit, it may be preferable to use oligonucleotides that are only 7-20 nucleotides in length. The microarray or detection kit may contain oligonucleotides that cover the known 5′, or 3′, sequence, sequential oligonucleotides which cover the full length sequence; or unique oligonucleotides selected from particular areas along the length of the sequence. Polynucleotides used in the microarray or detection kit may be oligonucleotides that are specific to a gene or genes of interest.

[0155] In order to produce oligonucleotides to a known sequence for a microarray or detection kit, the gene(s) of interest (or an ORF identified from the contigs of the present invention) is typically examined using a computer algorithm which starts at the 5′ or at the 3′ end of the nucleotide sequence. Typical algorithms will then identify oligomers of defined length that are unique to the gene, have a GC content within a range suitable for hybridization, and lack predicted secondary structure that may interfere with hybridization. In certain situations it may be appropriate to use pairs of oligonucleotides on a microarray or detection kit. The “pairs” will be identical, except for one nucleotide that preferably is located in the center of the sequence. The second oligonucleotide in the pair (mismatched by one) serves as a control. The number of oligonucleotide pairs may range from two to one million. The oligomers are synthesized at designated areas on a substrate using a light-directed chemical process. The substrate may be paper, nylon or other type of membrane, filter, chip, glass slide or any other suitable solid support.

[0156] In another aspect, an oligonucleotide may be synthesized on the surface of the substrate by using a chemical coupling procedure and an ink jet application apparatus, as described in PCT application WO95/251116 (Baldeschweiler et al.) which is incorporated herein in its entirety by reference. In another aspect, a “gridded” array analogous to a dot (or slot) blot may be used to arrange and link cDNA fragments or oligonucleotides to the surface of a substrate using a vacuum system, thermal, UV, mechanical or chemical bonding procedures. An array, such as those described above, may be produced by hand or by using available devices (slot blot or dot blot apparatus), materials (any suitable solid support), and machines (including robotic instruments), and may contain 8, 24, 96, 384, 1536, 6144 or more oligonucleotides, or any other number between two and one million which lends itself to the efficient use of commercially available instrumentation.

[0157] In order to conduct sample analysis using a microarray or detection kit, the RNA or DNA from a biological sample is made into hybridization probes. The mRNA is isolated, and cDNA is produced and used as a template to make antisense RNA (aRNA). The aRNA is amplified in the presence of fluorescent nucleotides, and labeled probes are incubated with the microarray or detection kit so that the probe sequences hybridize to complementary oligonucleotides of the microarray or detection kit. Incubation conditions are adjusted so that hybridization occurs with precise complementary matches or with various degrees of less complementarity. After removal of nonhybridized probes, a scanner is used to determine the levels and patterns of fluorescence. The scanned images are examined to determine degree of complementarity and the relative abundance of each oligonucleotide sequence on the microarray or detection kit. The biological samples may be obtained from any bodily fluids (such as blood, urine, saliva, phlegm, gastric juices, etc.), cultured cells, biopsies, or other tissue preparations. A detection system may be used to measure the absence, presence, and amount of hybridization for all of the distinct sequences simultaneously. This data may be used for large-scale correlation studies on the sequences, expression patterns, mutations, variants, or polymorphisms among samples.

[0158] Using such arrays, the present invention provides methods to identify the expression of the secreted proteins/peptides of the present invention. In detail, such methods comprise incubating a test sample with one or more nucleic acid molecules and assaying for binding of the nucleic acid molecule with components within the test sample. Such assays will typically involve arrays comprising many genes, at least one of which is a gene of the present invention and or alleles of the secreted protein gene of the present invention. FIG. 3 provides information on SNPs that have been found at 24 different nucleotide positions in the gene encoding the nephronectin protein of the present invention. Amongst the SNP variations was a non-synonymous coding SNP identified at nucleotide position 47,666 (protein position 281). The change in the amino acid sequence caused by this SNP is indicated in FIG. 3 and can readily be determined using the universal genetic code and the protein sequence provided in FIG. 2 as a reference.

[0159] Conditions for incubating a nucleic acid molecule with a test sample vary. Incubation conditions depend on the format employed in the assay, the detection methods employed, and the type and nature of the nucleic acid molecule used in the assay. One skilled in the art will recognize that any one of the commonly available hybridization, amplification or array assay formats can readily be adapted to employ the novel fragments of the Human genome disclosed herein. Examples of such assays can be found in Chard, T, An Introduction to Radioimmunoassay and Related Techniques, Elsevier Science Publishers, Amsterdam, The Netherlands (1986); Bullock, G. R. et al., Techniques in Immunocytochemistry, Academic Press, Orlando, Fla. Vol. 1 (1982), Vol. 2 (1983), Vol. 3 (1985); Tijssen, P., Practice and Theory of Enzyme Immunoassays: Laboratory Techniques in Biochemistry and Molecular Biology, Elsevier Science Publishers, Amsterdam, The Netherlands (1985).

[0160] The test samples of the present invention include cells, protein or membrane extracts of cells. The test sample used in the above-described method will vary based on the assay format, nature of the detection method and the tissues, cells or extracts used as the sample to be assayed. Methods for preparing nucleic acid extracts or of cells are well known in the art and can be readily be adapted in order to obtain a sample that is compatible with the system utilized.

[0161] In another embodiment of the present invention, kits are provided which contain the necessary reagents to carry out the assays of the present invention.

[0162] Specifically, the invention provides a compartmentalized kit to receive, in close confinement, one or more containers which comprises: (a) a first container comprising one of the nucleic acid molecules that can bind to a fragment of the Human genome disclosed herein; and (b) one or more other containers comprising one or more of the following: wash reagents, reagents capable of detecting presence of a bound nucleic acid.

[0163] In detail, a compartmentalized kit includes any kit in which reagents are contained in separate containers. Such containers include small glass containers, plastic containers, strips of plastic, glass or paper, or arraying material such as silica. Such containers allows one to efficiently transfer reagents from one compartment to another compartment such that the samples and reagents are not cross-contaminated, and the agents or solutions of each container can be added in a quantitative fashion from one compartment to another. Such containers will include a container which will accept the test sample, a container which contains the nucleic acid probe, containers which contain wash reagents (such as phosphate buffered saline, Tris-buffers, etc.), and containers which contain the reagents used to detect the bound probe. One skilled in the art will readily recognize that the previously unidentified secreted protein gene of the present invention can be routinely identified using the sequence information disclosed herein can be readily incorporated into one of the established kit formats which are well known in the art, particularly expression arrays.

[0164] Vectors/Host Cells

[0165] The invention also provides vectors containing the nucleic acid molecules described herein. The term “vector” refers to a vehicle, preferably a nucleic acid molecule, which can transport the nucleic acid molecules. When the vector is a nucleic acid molecule, the nucleic acid molecules are covalently linked to the vector nucleic acid. With this aspect of the invention, the vector includes a plasmid, single or double stranded phage, a single or double stranded RNA or DNA viral vector, or artificial chromosome, such as a BAC, PAC, YAC, OR MAC.

[0166] A vector can be maintained in the host cell as an extrachromosomal element where it replicates and produces additional copies of the nucleic acid molecules. Alternatively, the vector may integrate into the host cell genome and produce additional copies of the nucleic acid molecules when the host cell replicates.

[0167] The invention provides vectors for the maintenance (cloning vectors) or vectors for expression (expression vectors) of the nucleic acid molecules. The vectors can function in prokaryotic or eukaryotic cells or in both (shuttle vectors).

[0168] Expression vectors contain cis-acting regulatory regions that are operably linked in the vector to the nucleic acid molecules such that transcription of the nucleic acid molecules is allowed in a host cell. The nucleic acid molecules can be introduced into the host cell with a separate nucleic acid molecule capable of affecting transcription. Thus, the second nucleic acid molecule may provide a trans-acting factor interacting with the cis-regulatory control region to allow transcription of the nucleic acid molecules from the vector. Alternatively, a trans-acting factor may be supplied by the host cell. Finally, a trans-acting factor can be produced from the vector itself. It is understood, however, that in some embodiments, transcription and/or translation of the nucleic acid molecules can occur in a cell-free system.

[0169] The regulatory sequence to which the nucleic acid molecules described herein can be operably linked include promoters for directing mRNA transcription. These include, but are not limited to, the left promoter from bacteriophage λ, the lac, TRP, and TAC promoters from E coli, the early and late promoters from SV40, the CMV immediate early promoter, the adenovirus early and late promoters, and retrovirus long-terminal repeats.

[0170] In addition to control regions that promote transcription, expression vectors may also include regions that modulate transcription, such as repressor binding sites and enhancers. Examples include the SV40 enhancer, the cytomegalovirus immediate early enhancer, polyoma enhancer, adenovirus enhancers, and retrovirus LTR enhancers.

[0171] In addition to containing sites for transcription initiation and control, expression vectors can also contain sequences necessary for transcription termination and, in the transcribed region a ribosome binding site for translation. Other regulatory control elements for expression include initiation and termination codons as well as polyadenylation signals. The person of ordinary skill in the art would be aware of the numerous regulatory sequences that are useful in expression vectors. Such regulatory sequences are described, for example, in Sambrook et al., Molecular Cloning: A Laboratory Manual. 2nd. ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., (1989).

[0172] A variety of expression vectors can be used to express a nucleic acid molecule. Such vectors include chromosomal, episomal, and virus-derived vectors, for example vectors derived from bacterial plasmids, from bacteriophage, from yeast episomes, from yeast chromosomal elements, including yeast artificial chromosomes, from viruses such as baculoviruses, papovaviruses such as SV40, Vaccinia viruses, adenoviruses, poxviruses, pseudorabies viruses, and retroviruses. Vectors may also be derived from combinations of these sources such as those derived from plasmid and bacteriophage genetic elements, e.g. cosmids and phagemids. Appropriate cloning and expression vectors for prokaryotic and eukaryotic hosts are described in Sambrook et al., Molecular Cloning: A Laboratory Manual. 2nd. ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., (1989).

[0173] The regulatory sequence may provide constitutive expression in one or more host cells (i.e. tissue specific) or may provide for inducible expression in one or more cell types such as by temperature, nutrient additive, or exogenous factor such as a hormone or other ligand. A variety of vectors providing for constitutive and inducible expression in prokaryotic and eukaryotic hosts are well known to those of ordinary skill in the art.

[0174] The nucleic acid molecules can be inserted into the vector nucleic acid by well-known methodology. Generally, the DNA sequence that will ultimately be expressed is joined to an expression vector by cleaving the DNA sequence and the expression vector with one or more restriction enzymes and then ligating the fragments together. Procedures for restriction enzyme digestion and ligation are well known to those of ordinary skill in the art.

[0175] The vector containing the appropriate nucleic acid molecule can be introduced into an appropriate host cell for propagation or expression using well-known techniques. Bacterial cells include, but are not limited to, E. coli, Streptomyces, and Salmonella typhimurium. Eukaryotic cells include, but are not limited to, yeast, insect cells such as Drosophila, animal cells such as COS and CHO cells, and plant cells.

[0176] As described herein, it may be desirable to express the peptide as a fusion protein. Accordingly, the invention provides fusion vectors that allow for the production of the peptides. Fusion vectors can increase the expression of a recombinant protein, increase the solubility of the recombinant protein, and aid in the purification of the protein by acting for example as a ligand for affinity purification. A proteolytic cleavage site may be introduced at the junction of the fusion moiety so that the desired peptide can ultimately be separated from the fusion moiety. Proteolytic enzymes include, but are not limited to, factor Xa, thrombin, and enterokinase. Typical fusion expression vectors include pGEX (Smith et al, Gene 67:31-40 (1988)), pMAL (New England Biolabs, Beverly, Mass.) and pRIT5 (Pharmacia, Piscataway, N.J.) which fuse glutathione S-transferase (GST), maltose E binding protein, or protein A, respectively, to the target recombinant protein. Examples of suitable inducible non-fusion E coli expression vectors include pTrc (Amann et al., Gene 69:301-315 (1988)) and pET 11d (Studier et al., Gene Expression Technology: Methods in Enzymology 185:60-89 (1990)).

[0177] Recombinant protein expression can be maximized in host bacteria by providing a genetic background wherein the host cell has an impaired capacity to proteolytically cleave the recombinant protein. (Gottesman, S., Gene Expression Technology: Methods in Enzymology 185, Academic Press, San Diego, Calif. (1990) 119-128). Alternatively, the sequence of the nucleic acid molecule of interest can be altered to provide preferential codon usage for a specific host cell, for example E. coli. (Wada et al., Nucleic Acids Res. 20:2111-2118 (1992)).

[0178] The nucleic acid molecules can also be expressed by expression vectors that are operative in yeast. Examples of vectors for expression in yeast e.g., S. cerevisiae include pYepSec1 (Baldari, et al., EMBO J. 6:229-234 (1987)), pMFa (Kurjan et al., Cell 30:933-943(1982)), pJRY88 (Schultz et al., Gene 54:113-123 (1987)), and pYES2 (Invitrogen Corporation, San Diego, Calif.).

[0179] The nucleic acid molecules can also be expressed in insect cells using, for example, baculovirus expression vectors. Baculovirus vectors available for expression of proteins in cultured insect cells (e.g., Sf9 cells) include the pAc series (Smith et al., Mol. Cell Biol. 3:2156-2165 (1983)) and the pVL series (Lucklow et al., Virology 170:31-39 (1989)).

[0180] In certain embodiments of the invention, the nucleic acid molecules described herein are expressed in mammalian cells using mammalian expression vectors. Examples of mammalian expression vectors include pCDM8 (Seed, B. Nature 329:840(1987)) and pMT2PC (Kaufman et al., EMBO J. 6:187-195 (1987)).

[0181] The expression vectors listed herein are provided by way of example only of the well-known vectors available to those of ordinary skill in the art that would be useful to express the nucleic acid molecules. The person of ordinary skill in the art would be aware of other vectors suitable for maintenance propagation or expression of the nucleic acid molecules described herein. These are found for example in Sambrook, J., Fritsh, E. F., and Maniatis, T. Molecular Cloning: A Laboratory Manual. 2nd, ed., Cold Spring Harbor Laboratory, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1989.

[0182] The invention also encompasses vectors in which the nucleic acid sequences described herein are cloned into the vector in reverse orientation, but operably linked to a regulatory sequence that permits transcription of antisense RNA. Thus, an antisense transcript can be produced to all, or to a portion, of the nucleic acid molecule sequences described herein, including both coding and non-coding regions. Expression of this antisense RNA is subject to each of the parameters described above in relation to expression of the sense RNA (regulatory sequences, constitutive or inducible expression, tissue-specific expression).

[0183] The invention also relates to recombinant host cells containing the vectors described herein. Host cells therefore include prokaryotic cells, lower eukaryotic cells such as yeast, other eukaryotic cells such as insect cells, and higher eukaryotic cells such as mammalian cells.

[0184] The recombinant host cells are prepared by introducing the vector constructs described herein into the cells by techniques readily available to the person of ordinary skill in the art. These include, but are not limited to, calcium phosphate transfection, DEAE-dextran-mediated transfection, cationic lipid-mediated transfection, electroporation, transduction, infection, lipofection, and other techniques such as those found in Sambrook, et al. (Molecular Cloning: A Laboratory Manual. 2nd, ed., Cold Spring Harbor Laboratory, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1989).

[0185] Host cells can contain more than one vector. Thus, different nucleotide sequences can be introduced on different vectors of the same cell. Similarly, the nucleic acid molecules can be introduced either alone or with other nucleic acid molecules that are not related to the nucleic acid molecules such as those providing trans-acting factors for expression vectors. When more than one vector is introduced into a cell, the vectors can be introduced independently, co-introduced or joined to the nucleic acid molecule vector.

[0186] In the case of bacteriophage and viral vectors, these can be introduced into cells as packaged or encapsulated virus by standard procedures for infection and transduction. Viral vectors can be replication-competent or replication-defective. In the case in which viral replication is defective, replication will occur in host cells providing functions that complement the defects.

[0187] Vectors generally include selectable markers that enable the selection of the subpopulation of cells that contain the recombinant vector constructs. The marker can be contained in the same vector that contains the nucleic acid molecules described herein or may be on a separate vector. Markers include tetracycline or ampicillin-resistance genes for prokaryotic host cells and dihydrofolate reductase or neomycin resistance for eukaryotic host cells. However, any marker that provides selection for a phenotypic trait will be effective.

[0188] While the mature proteins can be produced in bacteria, yeast, mammalian cells, and other cells under the control of the appropriate regulatory sequences, cell-free transcription and translation systems can also be used to produce these proteins using RNA derived from the DNA constructs described herein.

[0189] Where secretion of the peptide is desired, which is difficult to achieve with multi-transmembrane domain containing proteins such as kinases, appropriate secretion signals are incorporated into the vector. The signal sequence can be endogenous to the peptides or heterologous to these peptides.

[0190] Where the peptide is not secreted into the medium, which is typically the case with kinases, the protein can be isolated from the host cell by standard disruption procedures, including freeze thaw, sonication, mechanical disruption, use of lysing agents and the like. The peptide can then be recovered and purified by well-known purification methods including ammonium sulfate precipitation, acid extraction, anion or cationic exchange chromatography, phosphocellulose chromatography, hydrophobic-interaction chromatography, affinity chromatography, hydroxylapatite chromatography, lectin chromatography, or high performance liquid chromatography.

[0191] It is also understood that depending upon the host cell in recombinant production of the peptides described herein, the peptides can have various glycosylation patterns, depending upon the cell, or maybe non-glycosylated as when produced in bacteria. In addition, the peptides may include an initial modified methionine in some cases as a result of a host-mediated process.

[0192] Uses of Vectors and Host Cells

[0193] The recombinant host cells expressing the peptides described herein have a variety of uses. First, the cells are useful for producing a secreted protein or peptide that can be further purified to produce desired amounts of secreted protein or fragments. Thus, host cells containing expression vectors are useful for peptide production.

[0194] Host cells are also useful for conducting cell-based assays involving the secreted protein or secreted protein fragments, such as those described above as well as other formats known in the art. Thus, a recombinant host cell expressing a native secreted protein is useful for assaying compounds that stimulate or inhibit secreted protein function.

[0195] Host cells are also useful for identifying secreted protein mutants in which these functions are affected. If the mutants naturally occur and give rise to a pathology, host cells containing the mutations are useful to assay compounds that have a desired effect on the mutant secreted protein (for example, stimulating or inhibiting function) which may not be indicated by their effect on the native secreted protein.

[0196] Genetically engineered host cells can be further used to produce non-human transgenic animals. A transgenic animal is preferably a mammal, for example a rodent, such as a rat or mouse, in which one or more of the cells of the animal include a transgene. A transgene is exogenous DNA which is integrated into the genome of a cell from which a transgenic animal develops and which remains in the genome of the mature animal in one or more cell types or tissues of the transgenic animal. These animals are useful for studying the function of a secreted protein and identifying and evaluating modulators of secreted protein activity. Other examples of transgenic animals include non-human primates, sheep, dogs, cows, goats, chickens, and amphibians.

[0197] A transgenic animal can be produced by introducing nucleic acid into the male pronuclei of a fertilized oocyte, e.g., by microinjection, retroviral infection, and allowing the oocyte to develop in a pseudopregnant female foster animal. Any of the secreted protein nucleotide sequences can be introduced as a transgene into the genome of a non-human animal, such as a mouse.

[0198] Any of the regulatory or other sequences useful in expression vectors can form part of the transgenic sequence. This includes intronic sequences and polyadenylation signals, if not already included. A tissue-specific regulatory sequence(s) can be operably linked to the transgene to direct expression of the secreted protein to particular cells.

[0199] Methods for generating transgenic animals via embryo manipulation and microinjection, particularly animals such as mice, have become conventional in the art and are described, for example, in U.S. Pat. Nos. 4,736,866 and 4,870,009, both by Leder et al., U.S. Pat. No. 4,873,191 by Wagner et al and in Hogan, B., Manipulating the Mouse Embryo, (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1986). Similar methods are used for production of other transgenic animals. A transgenic founder animal can be identified based upon the presence of the transgene in its genome and/or expression of transgenic mRNA in tissues or cells of the animals. A transgenic founder animal can then be used to breed additional animals carrying the transgene. Moreover, transgenic animals carrying a transgene can further be bred to other transgenic animals carrying other transgenes. A transgenic animal also includes animals in which the entire animal or tissues in the animal have been produced using the homologously recombinant host cells described herein.

[0200] In another embodiment, transgenic non-human animals can be produced which contain selected systems that allow for regulated expression of the transgene. One example of such a system is the cre/loxP recombinase system of bacteriophage PI. For a description of the cre/loxP recombinase system, see, e.g., Lakso et al. PNAS 89:6232-6236 (1992). Another example of a recombinase system is the FLP recombinase system of S. cerevisiae (O'Gorman et al. Science 251:1351-1355 (1991). If a cre/loxP recombinase system is used to regulate expression of the transgene, animals containing transgenes encoding both the Cre recombinase and a selected protein is required. Such animals can be provided through the construction of “double” transgenic animals, e.g., by mating two transgenic animals, one containing a transgene encoding a selected protein and the other containing a transgene encoding a recombinase.

[0201] Clones of the non-human transgenic animals described herein can also be produced according to the methods described in Wilmut, I. et al. Nature 385:810-813 (1997) and PCT International Publication Nos. WO 97/07668 and WO 97/07669. In brief, a cell, e.g., a somatic cell, from the transgenic animal can be isolated and induced to exit the growth cycle and enter G_(o) phase. The quiescent cell can then be fused, e.g., through the use of electrical pulses, to an enucleated oocyte from an animal of the same species from which the quiescent cell is isolated. The reconstructed oocyte is then cultured such that it develops to morula or blastocyst and then transferred to pseudopregnant female foster animal. The offspring born of this female foster animal will be a clone of the animal from which the cell, e.g., the somatic cell, is isolated.

[0202] Transgenic animals containing recombinant cells that express the peptides described herein are useful to conduct the assays described herein in an in vivo context. Accordingly, the various physiological factors that are present in vivo and that could effect substrate binding, secreted protein activation, and signal transduction, may not be evident from in vitro cell-free or cell-based assays. Accordingly, it is useful to provide non-human transgenic animals to assay in vivo secreted protein function, including substrate interaction, the effect of specific mutant secreted proteins on secreted protein function and substrate interaction, and the effect of chimeric secreted proteins. It is also possible to assess the effect of null mutations, that is, mutations that substantially or completely eliminate one or more secreted protein functions.

[0203] All publications and patents mentioned in the above specification are herein incorporated by reference. Various modifications and variations of the described method and system of the invention will be apparent to those skilled in the art without departing from the scope and spirit of the invention. Although the invention has been described in connection with specific preferred embodiments, it should be understood that the invention as claimed should not be unduly limited to such specific embodiments. Indeed, various modifications of the above-described modes for carrying out the invention which are obvious to those skilled in the field of molecular biology or related fields are intended to be within the scope of the following claims.

1 6 1 1839 DNA Homo sapiens 1 atggattttc tcctggcgct ggtgctggta tcctcgctct acctgcaggc ggccgccgag 60 ttcgacggga ggtggcccag gcaaatagtg tcatcgattg gcctatgtcg ttatggtggg 120 aggattgact gctgctgggg ctgggctcgc cagtcttggg gacagtgtca gcctttctac 180 gtcttaaggc agagaatagc caggataagg tgccagctca aagctgtgtg ccaaccacga 240 tgcaaacatg gtgaatgtat cgggccaaac aagtgcaagt gtcatcctgg ttatgctgga 300 aaaacctgta atcaagacga gcacatccca gctcctcttg accaaggcag tgaacagcct 360 cttttccaac ccctggatca ccaagccaca agtttgcctt caagggatct aaatgagtgt 420 ggcctgaagc cccggccctg taagcacagg tgcatgaaca cttacggcag ctacaagtgc 480 tactgtctca acggatatat gctcatgccg gatggttcct gctcaagtgc cctgacctgc 540 tccatggcaa actgtcagta tggctgtgat gttgttaaag gacaaatacg gtgccagtgc 600 ccatcccctg gcctgcagct ggctcctgat gggaggacct gtgtagatgt tgatgaatgt 660 gctacaggaa gagcctcctg ccctagattt aggcaatgtg tcaacacttt tgggagctac 720 atctgcaagt gtcataaagg cttcgatctc atgtatattg gaggcaaata tcaatgtcat 780 gacatagacg aatgctcact tggtcagtat cagtgcagca gctttgctcg atgttataac 840 gtacgtgggt cctacaagtg caaatgtaaa gaaggatacc agggtgatgg actgacttgt 900 gtgtatatcc caaaagttat gattgaacct tcaggtccaa ttcatgtacc aaagggaaat 960 ggtaccattt taaagggtga cacaggaaat aataattgga ttcctgatgt tggaagtact 1020 tggtggcctc cgaagacacc atatattcct cctatcatta ccaacaggcc tacttctaag 1080 ccaacaacaa gacctacacc aaagccaaca ccaattccta ctccaccacc accaccaccc 1140 ctgccaacag agctcagaac acctctacca cctacaaccc cagaaaggcc aaccaccgga 1200 ctgacaacta tagcaccagc tgccagtaca cctccaggag ggattacagt tgacaacagg 1260 gtacagacag accctcagaa acccagagga gatgtgttca ttccacggca accttcaaat 1320 gacttgtttg aaatatttga aatagaaaga ggagtcagtg cagacgatga agcaaaggat 1380 gatccaggtg ttctggtaca cagttgtaat tttgaccatg gactttgtgg atggatcagg 1440 gagaaagaca atgacttgca ctgggaacca atcagggacc cagcaggtgg acaatatctg 1500 acagtgtcgg cagccaaagc cccaggggga aaagctgcac gcttggtgct acctctcggc 1560 cgcctcatgc attcagggga cctgtgcctg tcattcaggc acaaggtgac ggggctgcac 1620 tctggcacac tccaggtgtt tgtgagaaaa cacggtgccc acggagcagc cctgtgggga 1680 agaaatggtg gccatggctg gaggcaaaca cagatcacct tgcgaggggc tgacatcaag 1740 agcgtcgtct tcaaaggtga aaaaaggcgt ggtcacactg gggagattgg attagatgat 1800 gtgagcttga aaaaaggcca ctgctctgaa gaacgctaa 1839 2 612 PRT Homo sapiens 2 Met Asp Phe Leu Leu Ala Leu Val Leu Val Ser Ser Leu Tyr Leu Gln 1 5 10 15 Ala Ala Ala Glu Phe Asp Gly Arg Trp Pro Arg Gln Ile Val Ser Ser 20 25 30 Ile Gly Leu Cys Arg Tyr Gly Gly Arg Ile Asp Cys Cys Trp Gly Trp 35 40 45 Ala Arg Gln Ser Trp Gly Gln Cys Gln Pro Phe Tyr Val Leu Arg Gln 50 55 60 Arg Ile Ala Arg Ile Arg Cys Gln Leu Lys Ala Val Cys Gln Pro Arg 65 70 75 80 Cys Lys His Gly Glu Cys Ile Gly Pro Asn Lys Cys Lys Cys His Pro 85 90 95 Gly Tyr Ala Gly Lys Thr Cys Asn Gln Asp Glu His Ile Pro Ala Pro 100 105 110 Leu Asp Gln Gly Ser Glu Gln Pro Leu Phe Gln Pro Leu Asp His Gln 115 120 125 Ala Thr Ser Leu Pro Ser Arg Asp Leu Asn Glu Cys Gly Leu Lys Pro 130 135 140 Arg Pro Cys Lys His Arg Cys Met Asn Thr Tyr Gly Ser Tyr Lys Cys 145 150 155 160 Tyr Cys Leu Asn Gly Tyr Met Leu Met Pro Asp Gly Ser Cys Ser Ser 165 170 175 Ala Leu Thr Cys Ser Met Ala Asn Cys Gln Tyr Gly Cys Asp Val Val 180 185 190 Lys Gly Gln Ile Arg Cys Gln Cys Pro Ser Pro Gly Leu Gln Leu Ala 195 200 205 Pro Asp Gly Arg Thr Cys Val Asp Val Asp Glu Cys Ala Thr Gly Arg 210 215 220 Ala Ser Cys Pro Arg Phe Arg Gln Cys Val Asn Thr Phe Gly Ser Tyr 225 230 235 240 Ile Cys Lys Cys His Lys Gly Phe Asp Leu Met Tyr Ile Gly Gly Lys 245 250 255 Tyr Gln Cys His Asp Ile Asp Glu Cys Ser Leu Gly Gln Tyr Gln Cys 260 265 270 Ser Ser Phe Ala Arg Cys Tyr Asn Val Arg Gly Ser Tyr Lys Cys Lys 275 280 285 Cys Lys Glu Gly Tyr Gln Gly Asp Gly Leu Thr Cys Val Tyr Ile Pro 290 295 300 Lys Val Met Ile Glu Pro Ser Gly Pro Ile His Val Pro Lys Gly Asn 305 310 315 320 Gly Thr Ile Leu Lys Gly Asp Thr Gly Asn Asn Asn Trp Ile Pro Asp 325 330 335 Val Gly Ser Thr Trp Trp Pro Pro Lys Thr Pro Tyr Ile Pro Pro Ile 340 345 350 Ile Thr Asn Arg Pro Thr Ser Lys Pro Thr Thr Arg Pro Thr Pro Lys 355 360 365 Pro Thr Pro Ile Pro Thr Pro Pro Pro Pro Pro Pro Leu Pro Thr Glu 370 375 380 Leu Arg Thr Pro Leu Pro Pro Thr Thr Pro Glu Arg Pro Thr Thr Gly 385 390 395 400 Leu Thr Thr Ile Ala Pro Ala Ala Ser Thr Pro Pro Gly Gly Ile Thr 405 410 415 Val Asp Asn Arg Val Gln Thr Asp Pro Gln Lys Pro Arg Gly Asp Val 420 425 430 Phe Ile Pro Arg Gln Pro Ser Asn Asp Leu Phe Glu Ile Phe Glu Ile 435 440 445 Glu Arg Gly Val Ser Ala Asp Asp Glu Ala Lys Asp Asp Pro Gly Val 450 455 460 Leu Val His Ser Cys Asn Phe Asp His Gly Leu Cys Gly Trp Ile Arg 465 470 475 480 Glu Lys Asp Asn Asp Leu His Trp Glu Pro Ile Arg Asp Pro Ala Gly 485 490 495 Gly Gln Tyr Leu Thr Val Ser Ala Ala Lys Ala Pro Gly Gly Lys Ala 500 505 510 Ala Arg Leu Val Leu Pro Leu Gly Arg Leu Met His Ser Gly Asp Leu 515 520 525 Cys Leu Ser Phe Arg His Lys Val Thr Gly Leu His Ser Gly Thr Leu 530 535 540 Gln Val Phe Val Arg Lys His Gly Ala His Gly Ala Ala Leu Trp Gly 545 550 555 560 Arg Asn Gly Gly His Gly Trp Arg Gln Thr Gln Ile Thr Leu Arg Gly 565 570 575 Ala Asp Ile Lys Ser Val Val Phe Lys Gly Glu Lys Arg Arg Gly His 580 585 590 Thr Gly Glu Ile Gly Leu Asp Asp Val Ser Leu Lys Lys Gly His Cys 595 600 605 Ser Glu Glu Arg 610 3 78785 DNA Homo sapiens misc_feature (1)...(78785) n = A,T,C or G 3 gagaaaattg agattactac ctgcaaggtg tcattacctg gtaagaagcc tatcaaaagt 60 ttgtcctcct gaaaaagtag ttattgctaa aagctagctg ttttgatctc attcttgctc 120 atttgttttt aagactgaga taatgaaatg tcactcccat ggcaactctg cctctttttc 180 ggaatgatca ttggtggtca tagttgcagc ataataacca gttagacctt ggaaatcctt 240 tagattctcc ttattccatg atttaacaaa gactgatata attagctaca ttttactgaa 300 gggagaagct aaagttcaca ggcagaattc aatttaatcc aatccatctg tttnnnnnnn 360 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 420 nnnnnnnnnn nnnnnnnnct attggttaga ctaatgtctc acgggaccca gagtggtagg 480 gaggcaatga caacacaata cattaagagc taccttagag catgctaagt gtaatagaag 540 tatataaaac actagtctca gccaatcaaa aagtcagaaa aggcttcctg ggggactgtc 600 ggctaaattt aaactcaaag ggtaagggga gattgtccaa atgaagaagg aagaggaatg 660 aagaggaggc taaatttaaa tgaagaagga gtgttttcta ggccaaagca aatatatgga 720 aaacaaaaga aagacgcatg tagatggggc attacacttt tccctccagt tatttatcct 780 gttttcatcc accactcttc gtcttttcct tagatctcca gttttttagc cgtatattac 840 cccctttctc tctaatcatc cattgcacac agtgaggttt atttgtaaat ctaacccagg 900 caccaactaa ccaaccaaca aacaaaaaca gctaaggagt agccactgga acctggaacc 960 atgcctacac ttacaaaaat ttgattttct gcagaaaaaa tattttgact cctacatttt 1020 tggacttcat tagaaggacc tgaaatggat gacaccaagc tgtttgccta aaataatgtc 1080 ccaagcctga attggcatgg atctttcttg agattaaaat agaaacttgt tttgctaact 1140 gaaaacaact tagaaatcaa agagccattt aagttgaaac cattattttt cctttccttg 1200 aagaaaattc ctgtttttac acacactgaa tgatcaggat agtgaatcac cctaccacag 1260 aactttccat taaaaatttg aagttgtaga aacctcaaaa agaaaatgaa gatgggggga 1320 aaacgtttgt aatgtagcaa atgannnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 1380 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 1440 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 1500 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 1560 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 1620 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 1680 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 1740 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 1800 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 1860 nnnnnnnnnn nnnnnnnnnn nnnnnagatg atcactgcac aaggctgtca ttcgaggtaa 1920 ggttgggaag aggggtgact tgattgggct catgtacatc ttaggataat catgttttca 1980 tttgaacaca gtcatttagt acaatacttt ttactctgca acaactgaac aatgattact 2040 taaaatggtt tctaagtgat cccctcaaga tcaggcacca caacactact aggcagtagc 2100 agcccttcct ctctggcagg cactcatccc tagaggggca gtcctgtcct aatgtgctcg 2160 ggagttgagc tgatagagac agaaacagtc tgctcatttc ttcttcaaaa cccgactgga 2220 gagccaaata tttcttttgg ttagttacaa aataaaaaat atgcctttgc caactctgcg 2280 ggaaaagcaa actcccagag tttgccttcg ctaatttgtc caagtcgtgc tgtggtttta 2340 gtaaatgcag acactgctca gctccagccc cataaacctc tctgctctag ggcttctccg 2400 ccctccggtg aaagctactg ctcgcccctg cagtcaccac ctgttcgggc ggaacctgcg 2460 gagcgtgcac ctacgcctcg ggctcctttc ctcctccact cccctttcct gctgggcacc 2520 ctgctttccc tctcccagag agggtttgca acttttctcc caggctgggg ctcgccctgc 2580 ttggctaacc cccaagagcc actgccgtcc cgcagcgccc ctgcccccga gttgcctgcc 2640 ccgctgggcc cccgggagga gcggagcgcg ctcacccttc gcccggggct gggagggcgg 2700 cgagtcgggc gcacgcgcac cccctgcccg cccctggcgc ccctccccgc gggcggtgca 2760 gctacccctg cagcgcctcc cctagctaga agggagcggg agggggctcc gggcgccgcg 2820 cagcagacct gctccggccg cgcgcctcgc cgctgtcctc cgggagcggc agcagtagcc 2880 cgggcggcga gggctggggg ttcctcgaga ctctcagagg ggcgcctccc atcggcgccc 2940 accaccccaa cctgttcctc gcgcgccact gcgctgcgcc ccaggacccg ctgcccaaca 3000 tggattttct cctggcgctg gtgctggtat cctcgctcta cctgcaggcg gccgccgagt 3060 tcgacgggag gtgagctggg ccccggggcg ccctctcctc cttcccgcgc taatttcaca 3120 ctcactgtct tgggtcactt ttccccgcgg ggtttcgtgg tcagagaggc gtctcctcca 3180 tccagaagtt gggccaccgc acagcgtggc gcgaggagag cggtccagcg gctccgagtg 3240 cccgcccgag gcggagaggg cgcgcccttg cgagtctggg accccatccg cggccccccg 3300 agggcgactc gccccggctc gggaattagg actgagggag aggagccgct ggagcctggg 3360 atctcggctc tgagggcgcg gtttagccac ctacgccgag gtgacgcgcg aaacatccct 3420 tacccgggaa actcccgcgc ctgaactaga cggctcttca ctggggaagc ttccaggccc 3480 ccggggggag gcccgggctc tgctcagggc tctcggggcc gctcacacag agagtgggtg 3540 cgagtcagcg actgggctac gggggagatt tgtgggcctc tccatttggt tttcttgagg 3600 gaaggagact caaaatgagg accggagggt gggcgctccg tgaatgtgag catgagtgtg 3660 tggatgtgtg tgtgagagcg cacacactgc gccgctcctc agactcgggc gagcctgacg 3720 gcggcgtgct gtgacaggtt ccaacaacct cgggccgcgt ctccgctgtc actcagccgg 3780 tcctcccgcg ctccggggcc gctccggtgt gtgagagaca ctgggtctgt cgggagggtg 3840 tgctcggtcc ccctcacctc tgtgcaatta cagactaggc tcgtcccggg tgcagatggc 3900 tgctgcgaaa agaggtttta ctctggcgca caccgtcgcc cggtcgcctt ccttcagcga 3960 cctctgcccc cacccctccg tgtaggtccc tgcctagtcc taaagaaaga tgccgcacct 4020 gttttacctt taatcttgga acgaatcaat tccacaattg attcgctttc ttgtacccgg 4080 aggcgaacgg aggggaacca gtggagcgcc aagaaaaagc acagctgctt gttcactacc 4140 tggtcggagg aaatagaaag ttcggggtta ttttgtactt tgggcctggg ggtagaaagg 4200 caggtaaaag aaaaggggaa ttgaaaaaga taaggagact ttaagaaaac gtgatagcag 4260 ccagagtgta gaccttttta ttttattttt aaaagcaatt ctgtgctcac atttgggtat 4320 gttatgcatt tcttgcacac atatttgcaa caagaaaccc atcacaagat atgcatatga 4380 tgtgaatgca tatagctttt tgtaacttta aaaagatgtc taaaacagca aattaaaatg 4440 ttgataccca gaggaaaaaa gtcatttagg tgtgactcta aacaaggaaa caatttagca 4500 aataatgtgt caacgtgtaa tcaggtggat aaatctgtat cctgaaatta ctttctttag 4560 gcattattta cattagagag gaaaatacta tgaattgttg atctaagcac gtttcaaacc 4620 acaaggatac ttgagatatc agctacactt taaactctcc ttggttttat ttagactttt 4680 ttctagtact tttttgtttt ttttgtttct gccacagtta tgtctcaaaa agagctctgt 4740 tacacttgat ttttgagaaa cctttctgct ccttccccca tccacctttt atatgaggca 4800 gaaatttttt ctgctttgac atgtcttact taatactttt cagtttatgg tgaatcagtc 4860 aaacctggct ttcaccccag taacaggtgt gggtctttga ggaatctgtt ttatttcaag 4920 cttcagaaac tatcctgtga gtggcagcct tgagaattgt tggtgttagc tggtgctatt 4980 tctacttaag aaagcatctt tccctcccgt tttttctcct acccttgtgt gttgggggat 5040 ggggtattta actgtctgaa atttgacatt agatcatagc agaactattt ctgtgaaggt 5100 gttttaacct taaatttctg caggcataaa agagtttgta gaaaattgtt gggtgtggtt 5160 gacatttttt ctgaagtaat aatacagagg aaattacctt tcttctcaaa agtgtcctta 5220 tttattttga atcttttttt ttggtaggtg gcccaggcaa atagtgtcat cgattggcct 5280 atgtcgttat ggtgggagga ttgactgctg ctggggctgg gctcgccagt cttggggaca 5340 gtgtcagcgt gagtatcaag cctggggact tcagttccct gggaggtgtg gctttccacc 5400 ttgttcatgg cttcacccca catatcagag ggttcattac tgagcaaggc ttggccttgc 5460 aggtctgact tggggatttt caggtacagt ccagactcct tattctgctt cttttcagct 5520 ttagccacct gtattacggc ccagctttgt cattcacaga gagccacctt aaatgtttct 5580 tttagcattt ttccccccaa atctgcatct ccttcctgtt ttcttaccag gtttattnnn 5640 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 5700 nnnnnnnnnn nnnnnnnnnn nnnnnnctga ctcgcttgct ccaggctggg agatggggat 5760 aagataatga gtctagaccg tatttacact ggggataaga taatgagtct agaccgtatt 5820 tacacaaaat aaactctgaa ccttagagta cttatataat cctcaccata gctgttgtgt 5880 aaagtggccc atttgtaaat ccttttttgg actggggttg tattgacagt gttttaaatg 5940 gaccacccag tataatgaaa caaagccact gaaacaacat tatttgtaag ttctataagt 6000 aaccaacctc atgtaataag taccaaatca gaaagctgat tttcaccttt tctgtgacca 6060 caaatggata ttttaatggt aaaatttaga gctacatcaa aagagtttga gaaatttagg 6120 aaatttgtcc ccagcatttt atcgcgatct taaaattgta tctcactcct actgcaaaaa 6180 aatagtcttc aaatgatcaa gtacctttcc agagcaccct tagagtgctt ggggcggggg 6240 tagggggcac ttctggtaag atgatgggaa ctaagttggg ttctacattg ggatatatat 6300 tttattgcta atgaggagga ggcttagagg aagagagaag ggcagttacg aaggctagag 6360 ctggcaatgg agaagcctgc cttagagatg ggttgctagt gtgaggagtc aggcaaattt 6420 aagttcagga aagttaggag ttccctctgc tattttaatt tttgaggatg cttgcaatgt 6480 cttccttaat tttgtgaaag agggacagtg acagtcacag attgactcta attgcacata 6540 aagaccacaa tctctggttg ggaatagaaa ggtaaaggaa atgaatgttt gcctacctgg 6600 tatggaattt gagaaccaac agattctaat aaccaaaatg tgaagaaagg acccttctgt 6660 tggcccaaca cacctacaca taaccctcct gagtgaaaaa tgagtagttc tatacctgca 6720 gtctccagct gtgcaaatac ttctgatact acagaagact aaattccacc aggcaccatt 6780 cttctttttt gatcatcttc ccttaaaaca atattgaata gactaaccag tgagtgtaca 6840 gcagctttgc ctacttcttt tgtttactgg aaactggagt taccaccatc tccctttaac 6900 agaatgtaat tgaccccccg tgcaaagggt ctagccaagc agctcgcaca tagcagttag 6960 gcaaacattc gtttgcttca ccttttatgt ttatacttcc aaaatcaaag tagttccaag 7020 gttttctatg ttacagtgaa ataaaatccc tttcaattaa aaaggcacaa atggttcttt 7080 actattaaca ttgaagttgg tgaggttgta aaaattagct caaaggtgaa tgtttccttc 7140 tgtgtatttt atttttccag catcttagat ggcgcaaatg tctcttgtca gttcagagtt 7200 ctgcctgtct ttgttttgat ataagcaggt agaggaatgt ggggctgaga agtaagcttg 7260 aaggggcaga acaaaccaaa aagaggctga tcagattgaa tgaaatatct ctgaaaactc 7320 ttgattattt taaagaaagt ctttatgaaa ttaaaagttt tgtcactacg tttgttcaag 7380 aaaatgcctt gctattgtat aacaattcaa tctaatatga ttccttataa agattccaaa 7440 gaacttctag cgaatttaat gtgagaaatg tttttgcctt ttcgaccttt agataatcat 7500 gtagttcttt cccataagga agggctattc tcccttcctc atcagaggtg ctctggttct 7560 cttcttttct gaatgattca ctttggaatt ttccttcaaa acagcatagc aaaacaaaaa 7620 gaaactattc ccattactgc atagatcttc ccaacttatc ccatagaagg tgtgtttgta 7680 ggtagggaaa ggtggtggta ccctcattat attatttaac agaccttatg ccttagtgca 7740 gtgacccttt ggtgagtgtt tattccgttt ggttgagcat ttgtgagatg tttgacttaa 7800 tattcatgtg agtcaaatgt acgtatctgt acaaaacatg ctgcccttca ttttcacttg 7860 ctatcttcct atccatggtc ttgtttggag aaaccgacta atgttgcagg atgctaaagc 7920 tggtagacct ctccttctgg ctcactatgt ctaagcagag ccagatatag ctgggaaact 7980 ttatatcctt ccctctagga ctcaggaagg aaggatcaga gatgctactc aaatgggcat 8040 agaacctgtc ctgctgcttc ctgcctgtac cacccaacat tctatcccaa cattcctgcc 8100 tgctagggaa agcatgaccc gttccagaaa taggcaggtt gtgttttcat agttcttttg 8160 taggtcatac ccttggttgn nnnnnnnnnn nnnnnnnnnn attctttgtt ttgtgttctt 8220 gactcttaaa cagatctcta gcatattgaa aattcaacat ttgattttct aactgtcatg 8280 ggctttactt ttattgacta ttgatgtggc tgtttattgt caggtgaaaa ttttttaata 8340 tgttcacaca ctgatgattg catatttgca gcacacagca tcttaaacca ctcagaggtt 8400 tgtcacaaaa tgtgtgtttc ttgctctgta atttttttgt cattttgatg gcatatttta 8460 attatgcttt tatttctctc cttctaccaa gtggagacct ctgtaacaaa gatttttcag 8520 ggttgcatat ttcatcttta aaagctgtgt acactgtgga aacaattgtt aaccaaaagc 8580 actagataat ttaggataac tgagttcaag ttctgacttt tgtacctgaa taggtgtcta 8640 acatttctca gctacagttt tcttattaca caagcatatt tccaaggtat gttgaagctc 8700 taaactgcaa ctgaaaactt ccttaatgta gaaaaactat ataggatcta aatattgtat 8760 ttttgtatgg ttgtccttct gtttactcgg agatttgact gtatatgtgg cttatgacaa 8820 tagcattttt gttaaaagca ttttatagaa gtgtgaagaa aaactaaaaa tacaaccagt 8880 tccaaggttt aacaaaacta ttccgtttct gagttctttg gctgtcattg agcaacttgt 8940 ggtttctgaa ggaaattatg tgaattagga tggttttgta tcatttatcc ttaagaacag 9000 ggaaaattga gatgttttct tatgtttctg ctggagattt tggaaagatg tgaaacctac 9060 acctacagat tgaccttgct tagttagctc tgaacctcct gctgcctctt ccacgtaaag 9120 tgaaaatttt ggattcttat cggcttcaga taaacttaca ggttagtgaa acatagggac 9180 tgagatatag taattcattc tgaagctgtt ttggagtggt caaataattt tagttggata 9240 atatatatta ctggctaatg attgtggata ttggaagtga tgaaaaaatt attgaattat 9300 ttctttctgc atttcaaatg aaaaggctat tagtttgagc agagaatttt gatttagtaa 9360 acaaaatatt taaatttcat gtttcatttc tttctcctat ctgggttcag atactcagtc 9420 ttataaatgg aacatgattt atttttgctc cctaaactgg ttattaactt cctgtccata 9480 atcacaaaac tatatagatt atatatttct ttgattattt ggattttgaa tactctcttt 9540 aaaataatca agagaaaatt agagctgtta gaatgttaga atttgttttg aaggccacac 9600 atagtgttcc ctccacagag aggttatact agtaaatgcc tttctatttg aggtcaacaa 9660 ctatgacaac ttccattgaa catgagttag tattttaaac gtaaagcaat ttttatacct 9720 gtatgcaccc aaaaagtaac aggggctctc aaaaggggtg gggattgtac tgtttacatg 9780 tatattgaag attgctagca gaattctggg gccagcttgg tggagcggag tacacttcat 9840 tgtccttagt gtagtagctt cctcctctaa ttttgaggtg agaatgcaga atctgttttt 9900 tgtttgtttg cttttaggta ggaataaaag caaggcaagg aataattttg attacttgca 9960 acattaaact tgaatccaca aatccttagg aagtgaagtt tttgattaag attatttaac 10020 tgccactttc cttgaaaggt tgtttaagaa catcatgtac ctttgggtaa cttcaagtgg 10080 tcttggaatg cagattccaa agtaagatca gcgttgaaga atcttgacct tttcaaacag 10140 gtaatttgtt agtatgtgta gtcttcaaag ttaagtttca gagaaattgc tttgctcttt 10200 tattcttttc cccaatcaga actgatcttt atgtaatact attaagatct actaatttcc 10260 tgaaatccct tcataagctt aatctggcca ggtcttaacc tttatggatt agaaatttta 10320 gtacttctta agctagaagg ccaggccaac taaagggaag cactctcctc tctgccagtt 10380 cagcaactag atctgtcctc aaatacctgc cacagggata tgctgcttga agttgcccac 10440 gcatgtacac actgggacca agaaggcact tctggtgcca gaaacaacac tgtgtttgct 10500 ttgtggaaat ttttgatatg ctttaaaaaa tgtaggtgtc ttctccctca cctcctgtgt 10560 gtaacttcca gcatttcttt ttgttgcttt ttatctacag aattcatgtt ctttgcattt 10620 tgagttagtt gaatccactg tatgctttcc agatgataat tagtgaagct caatgattct 10680 atgcagtgtc tttcagtggg taagggaaga tattcatcac aggggtgggc cttcatcaca 10740 ggggagatgg aagggcaagg gagagggatt ctgatggcct aaccctcatt attgcccccc 10800 actgccacat gcagacactc ttgccgcttt cccattgatt ttgtgcaata ttttaaattt 10860 ttccaccaat ctttggaaaa tattaattta ttttgtagga gaaaatgtat tcatttcctt 10920 aaatatccct cctcagagca acaagcatga ttagtttttg gtatacttta aaaaatattt 10980 tacctactta tgggtaaatt gcaatagatc tctcccctat cacccacttg ttttgtacaa 11040 atgaaagcat actatataca ctgctttgta tgctgctgct ttttcttaac tgtacatgtt 11100 gaaaggtttt atttatagac cttcttacnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 11160 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 11220 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 11280 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 11340 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 11400 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnc ctccttactt ttttatgcac tcnnnnnnnn 11460 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 11520 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn aaaaaacttt gaacatatgt taaagatgtt 11580 gttttgcatg ttcttagtgt ttctgcaaaa taaattttta tatgtggtca aatggcagat 11640 acatttgtaa tttttatcaa ttgcccaatg acttcttact cagtagagaa gatagtttaa 11700 ggggaaagac tgaatggcga tgctgaccca aaggctccaa caatcccata tccagagtct 11760 gatctttcct tgattttagg attctttgat cccttttctt tcccaagaaa tccctctgac 11820 aactcagtga atgtccatct gctcccatca tctgtttccc atcagaccag aggcaatggg 11880 ctgaagctag gagaatggag tgagatgtgt ccatttgcca gggtctccca gtggcttcct 11940 gcctatcatt tacttgtaag aaagagccac gcattccttt aggaattgct taattcattg 12000 atcttttatt atctttgtag tttagccata annnnnnnnn nnnnnnnnnn nnnnnnnnnn 12060 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 12120 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 12180 nnnnnnnnnn nnnnnnnnnn nntcaactga tggagttgat atccagagac aagatacaga 12240 gattctgtat cttgnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 12300 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 12360 nnnnnnnnnn nnnnnnnnnn nttagccata atttttatac ctttgtgtat agtatgggct 12420 tttttttttt ctagacttat aacctattaa ggaggaatat gtagaattct gagatctgaa 12480 taccattttg gggacaccgt ttatgataaa aagattttca ggaaatatta agacattttt 12540 gtaccaaaat actttttgta gtgtccttaa aaccataggg gcatatattt tctaatagtt 12600 attctaatcc ttattccaat tatatactta aattgtctcc ttttagaaaa gtaagaaatg 12660 catgattagg tgaaaaatta aaagacacat actaagagca agacagcagt gaaggtaagt 12720 atttctaact ttcctgtttt cctgacacct agttnnnnnn nnnnnnnnnn nnnnnnnnnn 12780 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 12840 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 12900 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 12960 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 13020 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnca 13080 tcatcactgt tgtatacctt tagatatgga ggacatatct aaagacattt gatttctgta 13140 ttagtatagt aattgaagag gtttcatttt attgctacaa attttcttat gtttaatctg 13200 taatgtaaag gagagtaaat catggctaat attacagagt agaaaccttc tgttcagtca 13260 tcttctattt ttgattcctt agaagatacc ttaaaagata aattaaattg atttctttat 13320 ttctatttgt ttatgcccta tcttgtttgg gaaagtatat aaagtggctt ccaggaattc 13380 acacaattaa aataaggaaa taggggctac atggaggaaa atgggggtaa aataaaaata 13440 ttaggaggag ggtttttcaa atgcagatat gtaagccata ggatcctaca tatgtcctac 13500 aatttggcta ctagcttctg gtagccaaag gtaaaagatg ttattgctga tttaatttgt 13560 gttgtttgtc agggaaaaag caaacaagtt atgtgggagg aattatggtt ttactgcatg 13620 tagttctgaa cgaagacttt cacgtggccc tttacggagt gggagtgaat gatgggatgg 13680 aaggggccac agcaacattc ctatcatgaa gccagtggtg gtttttgaaa agctgtttca 13740 tagaagatcc gtcaacataa gatgaggccc ttactctaaa agtacagttc acggaaagct 13800 gttctataca tagtttaaga agcattatgt gcatagcttt ctgacgttca gctagataca 13860 gcccagaata tctagctgtg atggatgaac tgtatagggt tttggatgga cagctctgga 13920 ttagacaaga tagtttcagg ttagaatcac taacaatgtt ctgaagtttg ctgtattatt 13980 taacagatta aaggccagtt cattttgtct ttctttttta agtcgatata ttttgaagat 14040 cagtaatcaa ctaatggagt tgatatccag agacattgtg aatagagcag ggatagggcc 14100 ccgcctaggc tccagtaaaa ggaggatctg caaagaaatt agtgggagat tttttaatat 14160 caattttgta tccattggat agcaatatat aaattcacca ccattttgct atatacatat 14220 tcatttccat gccagttatg gcttggaata aggaggaaag gcatgaacat tgttcccagg 14280 catcatttac cctactgatg ctgatacatg gatatgggcc agtgagccca ggaacaacag 14340 aatccctgac cagtgcttat ttccccaaag ttcctgattg ctataggtgt tccggggagt 14400 caaagtaacc tcaactgttt tcttaattca cctggattta taagggttga tttatataag 14460 ttgcttagac ctgaacagac tcaaagcaga gtctgtagga aatactctgc atatcaacat 14520 cccgtaccca aacctaagtc atcttttcac tggggtgtgc ggaagggctg attctcatgt 14580 actcttgaag ccctaggcaa tagaacctga aatcctgatg cacatatacc ctaggataat 14640 ttctctctca aaaaaaagca aatagtgatt ttaaaaatta tgactcaatg catattctga 14700 gagtgcagca atccagtaag tgtcacatct ccttgggagg aacaaagaga aggtcttcaa 14760 acgccttttc tgcagagact agcgtgaacc aagaatctcc ctgtctgaat tgtcaccgta 14820 tatcaacgtg gcagcgagct gagaaagtta gcattgccag ggccgaggtt ttctgtcttc 14880 acggtaacaa acaaatgatc agtcctcaaa gatagtaaaa tgtagccaag ataatttggc 14940 ttgaaaaaaa ttcaaagtga tgtcaataac cagctacaac cacagaaaaa tactagtagc 15000 atattgatgg tgcatccctg ggaacatgcc agggcaaatt ggggtgcaaa atacaccagt 15060 tatttaaaaa ttggtatgtt ataaggtaaa gaattcatta agtaatatca aatacaaata 15120 aactttttat tttgattcac taaaacttct ttttaagttt tctgatttta ttacttaatt 15180 actactgata cataattaaa gaattatggc cataatagaa ttcctaataa aatttctaat 15240 aaagccagaa tgaaggagta tattacaggc cagacatgat aaagcattat gatgtgtggt 15300 aaaatagtga catctatttt ttcattctgt attattttat aaattttctg gagaatttca 15360 gtttaaacag cctgctgaaa tactgttaaa tcaacctgtt attcttaact ctgatgggag 15420 gaaacagata tgaataataa aaatgatttc ttagctttag aatatagttg ttgctttgga 15480 gaacaaaccg ttttattcca attatttttt attaggagac ttcattttct gccatacatt 15540 agctttggta gatactagat gccaggggag tgcaaatttg agaataagga tttggcatgg 15600 gttatttgca tgttgagagt caagatttaa ctaaatttta aaaccaaaca cttcatttga 15660 tcaatatctt tttaaccatt ctgtagatta ataatataaa ttctccagag ctgacattac 15720 tttgctataa catcatcaga tcacaagatt agggttgcct tttgtagatg ttattcactt 15780 actctgaaat gttaggaaat atgtcatcac agtttaaatt tgtagtaata tatacaaaag 15840 gaaaacacaa ctaggaattt tggatttatg cttactttgc caaaaaccat gttgattttc 15900 aaaaaccttt agccnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 15960 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 16020 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 16080 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 16140 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 16200 nnnnnnctaa ccttattttc gtaaatcaaa gttcctaaat tcagtttcta aggaaagccc 16260 ctaagtggtc aaaggtgtgt gttgctgaat ttcagaaaca aactccatgg atggactttc 16320 cagacataaa cttcccacca aaatattgtt ccaaagtgtt tagtgcctcc tattttattt 16380 tagtgttaag attttgtagg tacttggtaa ttatcagcag aatatttaca taccaacaaa 16440 tagcaaagcc attgataatt aatagaaaaa caaaatcata gcttatgttt agttgtattt 16500 tttagtgtat ttcagtgttt agagttgatt ttatattgca tatactaact gtggtaatag 16560 tttctactgg atagattatt tcaagttcac atgattatag aagtatttta tgaagtttga 16620 catagataat ggtggtttca gggtcccttg tctcttgggt tgagttttta catcattgaa 16680 gaattaaaag attgtgtctt ggtgtctatg tatggccaag aattaagaca ttctgatgta 16740 cagtttcatt ttctaactaa ggcaaaaatg atttttgaaa caattacttt ttaggataaa 16800 agtataaaat tatgaagaaa taatgtgtag gtttgctaaa tgttgtgttc ttaaatcttg 16860 tgttgaggtc tgatgaattt ttttatatct ttaatagttc taaagtagtg tactgaaagt 16920 taggatcatc catgttgcta cttaacttgt tgtccatagg gtctgttgtc ctggatgttg 16980 gtgttataaa tgtacagagt taggtagttt tctgtgaaga gtttgcaggc taatatcact 17040 gtttttgaca atgaaaaata tagcatgaaa attaaggttg gggtatggag aactttctag 17100 agctattgct ttttctagct gatgattaag gttgagaggc agaagtacat tgtgtcaggg 17160 aggacctttc ccttttatgc atgcacctgg catttatcac ctttaacaaa gtgtgtgtgt 17220 catgctgtgc tgcttgctta aggggctgta tgcctccagt ctgagtcaca tggtaactgc 17280 atcagcagtc ttagcctgta gcattttatt atttcttttc aaagtttaca cttggcctaa 17340 tacttagaca ttttataatt ctttacagta aatgaaacta tacatgagaa gatgggcata 17400 cctttggatt aaaaaaaaaa aaggctcagt tctttaaagt ttcttatcct tgattttcct 17460 aacacggtcc aaagttcagg actggctcca aacccataaa cctgtgttta gcaagcaaga 17520 aacataatcc ccagatagtt tagtttctct ggtttatgag tcacaatttc ataaaatcat 17580 cagagtgctt attaattcca accacgtata gtaaagaacc ttcagatgaa ctgaagcaag 17640 ggttcttagg caagttgccg gagagagttc tagaattcta aactacctga gtagctttgc 17700 tgaaatgttg cttgtatttg ctactgtggc cattttatga tggccataga gcaacagatt 17760 atcaagagaa aatgagacag atttttctgt attatgtgct atgaatgaat cctattttag 17820 tgaatgtttt aatggggttt ataccgcaaa aaaaaaaaat tgtatgtaag gcatttattt 17880 ctggcaactt tcataaaaat tgttgtgatg gttgcatata aaaatttcct tatccttcaa 17940 tagaggatag ttcccagaaa cttcctagaa gtaatctatt ccagatttaa cattgctttg 18000 acataaaatg cagttttgtg tagttttaaa atgcaaatta aaaaatatag gacattggct 18060 aaaattttat cttgaagtcg ggtatatatt gataccataa aacttactag atctatgtat 18120 ttcaaggcta atttatgcca agtaggaaaa atatgaccca accttaagat attacaagga 18180 taaaatagac tatacaaaac tgtttggcta tttggtacta atacaactag ttagaacata 18240 atgatgtttg ctattcttta ttaagttgtt ttacctttgc ttacaataat ttaaagtatt 18300 tttcctgata aatttgatga ctcaaaattg gcaattaaag aatattaaag aaacggtatc 18360 cttttatatt ttttctgtct catatataac catagtcata acttgtgtga tccagaatgt 18420 aatttgctat ttacactttg atttcagctg tttgcttaga ttgtacctga tgtattttat 18480 tattctattt aaggaatgtg tcaacatcaa gatatggtga gttcttttca aaataacagt 18540 agaaacctga ccaatatgaa aaaaaaaagt ctagcaagca aatgtaattt gtgtctttaa 18600 aaatacatag caatcattct ggatcaatag ttaaattatt gcttcaatta aatcaaattg 18660 gacttagaat ttttttcttc ttatattacc caaaggaagg ccccattcac cnnnnnnnnn 18720 nnnnnnnnnn nnnnnnnnnn nnnnnnnnaa ttgtaaatta cagtggatat ttaatccttt 18780 aaaggcatta atttagcaga gaagaataaa attatccctt ttcttacctc taaaatctct 18840 aggttgatca aacactgacc attattagta ccatttcaag tttcttccct ttactttcat 18900 caaactgggt tatttatgta tgttcaagtg aaatggctgt gctttcatgg taattcttgt 18960 tgctattgac aaccaaagca gccatgcaag aagaaaatgc tgtggaaggg aagaaaaaaa 19020 ttatatttcc tccccaaagt tggagagaga agggaacata cagtatgtaa gaaacaggta 19080 agtaaaaata tatcaatttt aaaactatta gcctttctta ctaattgatc aaaagtttaa 19140 attttttcag atgtgttttc caactccaat ttaaataaaa ggatactgtc tctaggaatc 19200 agaaattaat attctggaaa tgaaaaattg gatttgaaaa tacagcatca caaaaggtct 19260 gaaatattta aatttagaat ttggactata gaagaatagc ataacttcaa ttattggtta 19320 atatttttgt tatgaaatgt ttttttataa caaaatggtg actaaaatat tactatttta 19380 aacatgtcct agattttttt ttgtttcaga aaagcactga aagttgaata tgtgtaagtc 19440 tccgggaatg taacaagttg ataaataccc aagtccagcc tccttacaga gaaaagatct 19500 ggaatttctt tttttggaca tttgttttgc agctttctac gtcttaaggc agagaatagc 19560 caggataagg tgccagctca aaggttagat gaacatattt cttgaaataa ttttggctaa 19620 tctatgtctt gaaaggcata ccttctaaat aattttacaa agctgtaaac aaaacattag 19680 ttgtgttttt gaattgcttc tttttaggaa aactatattt cttagagatg tgcttattct 19740 atacataatt atagtaagtt aattttaatt ccattattac attaaacttt tccgtttcag 19800 ttgattattc ctttcttctg ttttatgaat ctaataaatt gacctccatt ctaagctaaa 19860 ggacagttta tgtttttaat acttaaggaa gcaaatataa aaacaaagtc caccaaatga 19920 aattactaac accagcctat cattaacctg agggagttgt atttcatatt cattaattct 19980 ccatggaaca attcaggtaa gaaatatttt tattgtatga aatatgttat cagttataat 20040 agcacttacg ggcatttatg ctcttgtttt aggtacataa agggttcctt gtttaagata 20100 tttttggcag taaataatgt cacttatcac cctatgacac tttagtggaa aacgttggtg 20160 aaatatgtgg aattgtaatg tttaatggaa ttgattctaa gaagacaatg nnnnnnnnnn 20220 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 20280 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 20340 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 20400 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 20460 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nngagaatgt 20520 tctttaaact gattgaagaa actatcacat gtcatgaaaa atgtttttaa ggcagnnnnn 20580 nnnnnnnnnn nnnnnnnngg aacaaaattg atagtactag aaatagaagt tcagaaattt 20640 ctttgttcag ggagacttaa agcatatgta ttttaaagtt actagcagtt ataggattga 20700 ctagtagagc tatggcttac attagggagg caatgccaag gaaagaatag aatgtgtggt 20760 tggccaccaa catnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 20820 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnctaacttt gcaagattct 20880 tactggttat aaggaggtgg cttccacagt tcttgacaga tgtttgctat tattagtgtg 20940 ttctcaatat atacaataca aatggagaac tctgagccga gttttactag gatccatcct 21000 aggacagatg gtacagcaca ttttcaaatg gttttgttta ccataatcat cttttggaaa 21060 taatcaattg gaagtagaac aattcttcag ttattcattt ctaaaatcaa aactttcatg 21120 ggatagtttt ctgtgcttct gaaccgtttt tcccatttct ctaataatat ccatccataa 21180 acctccgagt ggaacagttt tggttgtcta acagtgtttg tgcttgggaa atgaattgaa 21240 gggagatgga attgaattgt tgaatgtgag catttagagt tcctgtaatt tatccgtnnn 21300 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 21360 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnntttg taagatgatt ggctcttatt 21420 ccttctatga ttcttaaata ggaaaaagtt agggataaag atttgaggga gttcagggaa 21480 agaatcattt cctgctgaag aggatcaagg aagacttcat gaactcaatg gtattcagtt 21540 gatactatat atatatatat tatcagtata tctagtatat agtatcagta ttgaaagccc 21600 acagaggaag gcaatactgg gtaagaggca tgttttagta taggattctg tgagagcata 21660 tcttaagggg cagtagccca gaccagcagt gatggagtgg tgatggccca gctgaatccc 21720 gagggataaa tcagtcaagt gaagagtatc ctgggcacac agaaaaatgc atttaaagct 21780 cagtagccag agggaggagc acattccgag aactgttaag aatctcaacc cagcatatgt 21840 ggggtatatg tttgaagggc atctgatgag agacgcagtg ggagaagtgg gaaaggactc 21900 ctggaaggct tttgatatca tattaaatac atttgcctta atcacaaagt tgtagaggaa 21960 cactgtaaag attttaagca gagaaattaa tattttagag gatagctggt ggtaatggat 22020 cagatctagc actcaggtga atttagtttt gcttggagtg tttgtagttg tcagtgtctc 22080 acaggagaca cggatataag ggttggttca aaaatcggta ggtctgcatt gctgcgagga 22140 gtctatcagc tggggcctct taagtagctg gcccatttgg aagaagtatg tgcctcattt 22200 ccctgtggag ctcacttcct ttagtgcaaa tgcctgggac ctgtaggaag tcaacctggt 22260 gagaaattag gcttcatttt aagtagtggt ggtgggaaca aagagaagga gagggattga 22320 aagatactga aaagaaagga tggagaggca cacagtgtta gggcagtggc gggggggcac 22380 atgggcttgg cagacagact tgnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 22440 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 22500 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 22560 nnnnnnnnnn nnnnnnnnnn nnncagtatt tgtaatgctg gttgtgaaag gtgaagaaga 22620 tgtaatcaag attactccca agtctctagg acatcatttg gtgagctaga gaacacagga 22680 gaaggtgcag ataatgattt caattttgga tatgttcagt ttgaagcgct tgtgagactt 22740 tgaagtagag aaacttgata tgtgcttgga ttgcagggtg agatctgaac agaacacagg 22800 ctatgatttc atttctttcc actattctac tcaaaccttc cttccttttc cttcgtcaac 22860 ttcagtcaac attgattatt gccacaacta tctccacgga ggaacagctt taaacagtag 22920 aaaaagaatg tgggataggt aaaaatctgt gttccacatc tgctaaatgt tatctaatag 22980 ttgtaagttc ttgaaaaaaa atctttccag tttaccgttt tggcaaagca ttgtaagcaa 23040 ctaataattc aacatttgtt tatatcaagg gctgaaaaaa agtcatcctc tacaaacata 23100 ttttcttttt ttccctcttt tggttgcctt tttttgtttt acatcaccac cactcctaac 23160 tctaagaaaa tatttgacta aaagtgaatc attgttagta gtgaattcgt acnnnnnnnn 23220 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 23280 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 23340 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 23400 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 23460 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnagcacat 23520 ttatttttta agtttatatt ctgtcctaat acaatgtatt aaaaatgagc cctgcctcaa 23580 gtcctagcct aaatgccact ttctccagtc tgctttcctg attttcacag caggaagtct 23640 tctgtacttc ctctacatat aacacttgca tgaagcatat ctcacattgc cctacattgt 23700 ggttatttat ggatctatat tatttgttct gctagactat tagtttcatg aaggtggagg 23760 cagtgtctta tacatcttta cgccagtata acagcttctg caatgcctcg tccagttcag 23820 ttcagtacac ttttgaaggt gtaagagcta gtcatttcta tttattaata aaagggattt 23880 aaaaaaaaga tggacaagat gtaatcttaa taaacatatt gtggttttca ggaaacagtt 23940 taatgagaaa atgaatatat ttgtctccct accttatgac cttattacat atcatagatc 24000 ttcactcttg ctttagattc taaacagaaa tcaatttatg tgcttgaaat cacataaagg 24060 taatctgatt gtgtcatcta ctgcttacag tccattggct gttcacaatt atgggtagaa 24120 tcttgattat ttgagtagaa gccttttcag gctgacattg ccttatcagt ggaggactgt 24180 ttcttgttgt aggcccagtc gcagtggacg gcccttgggt ttccagactt ctgctgcttc 24240 atgcctgttg gcttttctta tgctgctcag tctgagtcaa aggcccttca cccattccca 24300 tggtgaattt ctacttatcc tttaaaattc agctcacatt tcctcttgaa agtttttcct 24360 ggtatctttc ctattcctgc ctccctgcag agaaatgcct tcttctgtat tcttttaaca 24420 ccttgtacac ctgtcatgtc tatctttgtc aggagactgt acactacaga gcaagtactg 24480 tgtatttaaa gagctcaaag tctagtgaag gagctagata agaaaaacaa ctctgaaaat 24540 atggtataag atatatatat ttaaaagtaa acctgtatat acatataatt tatgatagaa 24600 gtatgcacag aggcttatga aagcactgaa gagggtatga actcagcatg aggcgtgaaa 24660 atcttggggg tgatataatg gtcaaactga caggtgaagc ctgtctagat attagtgtag 24720 cagacaagag ggggaatggc attccatgtg aaagtgtgga ggtactcgag agtgtatttc 24780 taagaactgc aattaactgc tatatttgtg aatgtgattg gaaatgaggc caatgggata 24840 ggcaagggtc atatcatgaa atatggtact gaatgccatg ctgagtgttt gaaatagtat 24900 aatgatctac atgaacatga tgaggagcca gtgaatgagt ataaggagca gaatggcatg 24960 ctgaaatcat gggttaggga ggtccctcta acaatagttc aggcctggat tgaagtccat 25020 gatgttggac acagaaagac tgtnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 25080 nnnnnnnnnn nnnnnnnnnn nnnnnnnnna aggttcttca acagataaat ttggaataat 25140 tatcattttc tttagaaaac aaataagcaa actaaaaaca cctgaaggtc aaaatgacaa 25200 atctctgaca tcatattagt cgaggcccag taacaaaatt gnnnnnnnnn nnnnnnnnnn 25260 nnnnntatac ttcaactcat gtatcagtga ctcttttaaa catagagatt ttggttgcta 25320 aaaaaggtga tgatggttac attaaaattt tctggccttt tgtgatgatt ctttagcaaa 25380 cctgctcaaa attacagtat cagatgtttt aaaaaatgat aattttcaac tgaaagaaat 25440 actgtaatgt atttcatcat attctatgaa ttcttacatt atgaaagata tcttttatgt 25500 ttcagatatc tattatattt cctttttgaa agaatttttt ctagcactac atcttttcca 25560 aacctttttg actttcctaa gcctgtgtaa ttaatcactg cctgttctgt gtccttaaca 25620 tgcatctgtt atcaggacat ctctgcttac ttctgtttct ccttactaga cttacctgct 25680 ggaggatgag aacttgctct cctattcatt tttatatccc caatttttaa tatagtgtct 25740 tagacactgg ggcgtttcct ccttgtcata cttataatcc tcnnnnnnnn nnnnnnnnnn 25800 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 25860 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 25920 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 25980 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnncctt 26040 tctctaaatt acattgaaaa gttaacaact actcacaagt taaataatag ttgtctgtgt 26100 attcatgtta ttgtactcct ttttatatat ttcttccatg nnnnnnnnnn nnnnnnnnnn 26160 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 26220 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 26280 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 26340 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 26400 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 26460 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 26520 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 26580 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 26640 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 26700 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 26760 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 26820 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 26880 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 26940 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 27000 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 27060 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 27120 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 27180 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 27240 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 27300 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 27360 nnnnnnnnnn nnnnnnntac tctactnnnn nnnnnnnnnn nnnnnnnnnn ttcacatttc 27420 caaataaata gtaacacttt ttaatgtgtt cttaacgttt tatttggcat taatctaaat 27480 tcccctctag cacaatgaaa acagaaagag aaagttaaaa tttcaagtaa actgaaaaca 27540 acaatagtgc tcagaggatt tttttttaag tgaaaaggga tagtgcttaa ttatgacaaa 27600 taaaagctaa cttgagatgc acatatacag atgcagccac ttattttggc gggggacact 27660 tcaggaatta aaatttaaat agcgagagat caaatagata cttggtaaat gtgtctgaat 27720 tggatgttcc cagacacaaa aataaaatga gttattgaca gctcttggga gacaacatta 27780 taaagactag acacgttatt tattttaact ctatgttcta aattaccatt gagtaattga 27840 cattcgtatt tgactatggt ttgtggttaa gttcttaatt gcaataatgt taaataaaat 27900 gtgaagccca aagcaaacaa caacaaaaat tatagcaata cttcaacaga ggtaataata 27960 atatgctgca tcaatggttc agaatccagc atctacataa aacaagcaac agggtaatga 28020 aattattttc ttttcaaata ttctggcaga gctactttag ttttcttaag ttatagattg 28080 tggtcttaac tgcaactttt cgctcctttt aagaagtatt taagttattt aaatgttact 28140 taaatacttt tatgttttta atcattatta acattctcta cccctctcat cctttcctgc 28200 ttagtatttt gttataatct cactctcccc acttccaaat gcactcaaaa atgctggact 28260 ttctggtctt tttctgacca ccagaagaag tagtggattg agcatggaac tgggataaga 28320 catttttccn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 28380 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 28440 nnnnnnnnnn nnnnnnnnnn tgagttctgt attcatgagg gcatttaatg tgtcttatag 28500 tgataacaat cagaggcatt attattcagt ttttactgca gcaggagttt agatttgacc 28560 atcagataat tgacactata gtacagggta ttaggaaagc tggaaagatg ctttcccctg 28620 gagatcttgg acagtaggat tcttctaggt tttgttctgc tcatgatatg caatggctca 28680 gggcactttt cagaggacca ctgtacaatc tccctccatt ccacccaggt cctattaata 28740 tagtagtgag ttgtttcata ttgccttcag tgactttccc ttttccaagg ccaattacca 28800 tttgaaaaag tcattccctg tcatattttc ttgttctgct caaatgaatt tgtccatatc 28860 tgaattctgt gaagcttgtt gggtgtaaaa acagcttttc aaagtcttcc agtttcatat 28920 ctatttgttc ttctgcccta cccttgcatg tttctctgtc cccttttctt ctgtgagcat 28980 aacctggagg acaaggtttt ttgttctgct tttagtggtg ccatgtctgc ataataacat 29040 gatgtagatt gaaaaaatta caaatgattc ttggaattct aaagataatc ttatttctat 29100 tgagaaaatc cttctcaagt tactaactac cattgagatt ggattggctt ttgccttatt 29160 acttttcaga gcctccatct gcttgggtat ctcaacatat ccttagtttt caaatgttgg 29220 cactttactc ccagcatgat taccttacat aagaaacatt ataggactga tgtgggagtt 29280 tacttttctc ataacttatt tgataattca ctgcttatgt tagagttaga aactattgtc 29340 caactctcag agacccagtt acatcactta agatggatan nnnnnnnnnn nnnnnnnnnn 29400 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 29460 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 29520 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 29580 nnnnnnnnnn nnnnnnggat ggatttttaa gactaaaatt ttagattgca atcaaacaga 29640 agaggcttgg acatgtgaca ttaaagagct atgtttgttt tgccagttga aacttgtttc 29700 tttttctagt taaaacaatg atttagaggt tattttgagg gcactttgaa gattatgcta 29760 caggaatgct agagagcaaa ttttgagagt gattgcccat ttggacttaa ttctggcaac 29820 tgattttggg gtaaaatgtc tatcattcgt tgattatctg tgatttttac ctggacttta 29880 cttaagcttt attaagttgc taaaccatat ttggatgcta gtgatagcag accatcaaat 29940 acggcccaaa cttcttgttc tgatccacac gaaggtcaga gaagcaatgc tgccttttct 30000 gatagccagt agcaccagca ggcacgttgt ttactccaaa caagaatttt aatatttttg 30060 aagaccactg aaaatggatc atttatactt ttttattttt ttgataagga agggatgacc 30120 tactattcac agagtaatgc agtttgctga aaaggttggt ttttgctgac ctctgagagc 30180 tcacattaca gtggagtgtg ttattggaag gtgagccagt ttatacagag gttgggaatt 30240 atttttctct agttttgaaa tggttatgca ctttctaatc tagttattta gatgtagaaa 30300 taaagttttt tttttacatt cccctgagta tatggtattg taggtataaa ataaatttga 30360 tgagttttat ttctgtacta ggatctactc ataactcttc tatcctaaat ttgtatcaaa 30420 aggaaacctt tgctgtcttg ataataaaca cagactaagt ccagattcca cagtctagtg 30480 atcaaggaaa ttcaggaatt gtatttagct acaagtaacg tgacacaaag aacagtgccc 30540 taaattgcta ggatggtgat ttagggttaa tattatgact tcttgctcta ccctctttcc 30600 attctcaaga tcgcctcatg gtcataaaga ggccactgtg gttgagtcat gacatgcacg 30660 ttctaggcga gaacacagaa caagccgtgc tcttcagccc ccttcttaca cagcattcca 30720 aagccccacc ccatcacttc tgctttcatc ccatttagcc agaacttagt catttggctg 30780 cccatatctg ctaaggagaa tagggaacat agttttccat tactaaccca tcccccactc 30840 nnnnnnnnnn nnnnnnnnnn nnccattcta tattaaagca gaagggaaag agatattggt 30900 aagaatccag ctggcctttt gtgatctgtg tcagcctttc ttttgatctc atctgctgtt 30960 ttaagcactt tacactgtag ccccacgaga acactttgca ctcactaaga agcagtcccc 31020 tttgctgcgc ccccccccca ccactttgct tatattctga aagtcttttg tttcctattc 31080 cactgctctt acctctaaca cactgcctct aacacaccaa cctgcagttg tagttattac 31140 acaccctcct tggttctttc atctctctat cacagccctt gttgtggttt agccaatata 31200 ttttagttcc acagctaaat tttcataccc tctatgactc tctaatcccc tgccacactt 31260 gcctactata atacattata tatataacaa atgtttgata cgtatttatt gaattccatt 31320 ccagaactaa tgccagcaag ataactttgt gctatatagg agaatatctt tttgtgcaac 31380 agtttccaaa gggttttctt tttctaagaa gaaagaaatt gattgtatca actttatgag 31440 tatcctaccg catttaatag ccattggcta atctaagggt tcctggttac ttcactgaat 31500 agcctatcag atggaagtgc aaacaacagt ttgttttgaa ataggactcc ctaaacatgg 31560 aagaaacatt aacagtgttg gcctgttgga atgtgtgcat ttgatgtgct caagattagg 31620 gcactctgct tgagaacaaa taacaaaaaa gggagaggaa acaataaaaa ctttggtcct 31680 ataaagcacc tgaaagtact ataaattgat ggttctcaag ctggtcaggg ggtccaaagg 31740 ctacagcctg ggggcctcaa gtttaattgt tttataaagt gtcctaaata aaatttttat 31800 gtttttaagt ggtattttta aaactacttt ttcatgcttt gagagagttt tccaaattcc 31860 aattatttta aggggttatt tcctggactt gcacttaacg attttgagat gtttacattt 31920 ttttcaatat ggcattctgt gtgcctcagt gatacatggt tatccaggtt gcatgcatat 31980 ataaatgtta agatttatgg aaggtcatct ttttagatta aaaagaattt ttttaagctg 32040 gtatttcttg gtgatagggc ctagaaatta tgtagagtgg cttacttctg gaacttattt 32100 taaactgcat ataaaccatc cgcctagtgt acagttggct aaagagtaat attagaaggc 32160 cctcctggac agtttatttc atttcatgga tatgaacaca attgtttccc tttgaattta 32220 atgccatgtt taaaatcaga ttttaagaat tttccaaggg catttcccta tcatttacac 32280 tctgcttgtg ttttctttct gtagtctttt acattaaata cctcctacag agcactgcct 32340 aaggatttgt ggtggtacag ggtccagttg ggatgacaaa caggcaagga aggcctggaa 32400 gtaaaattag caaagaggcc ctgtggaatg gaaggtgagg gaagggttag tgacagttgg 32460 gggaggaaag gtagaaaaaa aaataacatg cacatcagtt tcgcaggagt attagagtct 32520 taaaggaaac aatgtttgat aattatcaga gaggaaactg ggagcataga gcatatccca 32580 gaatggagaa cagcatgggt taaaatgggt aggaacaggt gtcaggagct tcaaannnnn 32640 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 32700 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 32760 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 32820 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 32880 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 32940 nnnnnnnnnn nnnnnnnnnn ntgattttta tcaggcagtt atgatgataa atgtatggaa 33000 acttcccatt tcctagagct aaagtgcatg tttctcattc tgaaatgtag ggaacataat 33060 catctgatac cactcacctg attgtttctc actcttcctc caccatttac ccatctcttt 33120 agcttaatga gtcccctgtg tatctcccaa ctaaacagcg gcttacttgc ctgtgaaata 33180 ttcttctctt gggtagtctg ctcccttctc tgtctactca tgcttcaaga ttcaacataa 33240 gcctcctcta tgaggctttc tgcacgtatg tatatggatt tgcttgtgta atgatttctt 33300 cacagatttc atattgctga taaataaata ttgttttgaa taagaaacgt ggttttgtat 33360 ttttatctcg attgtagact ccttgagacc agtaccatgc tatacaatta tttttcatct 33420 attatagtgt ctggcatagg gacatgcaca tatttggtac agaannnnnn nnnnnnnnnn 33480 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 33540 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 33600 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 33660 nnnnnnnnnn nnnnntagag aataaacgga ataaattcca attaaacagt gaaaatattt 33720 gagtgatgat ttacagaatt ttaaagtctt gagaaagtgg aactcagttg atggaaaaga 33780 gtgaatgtca aactgagaac gtcctatttg ctatgttagg gcatagaaag gccattatag 33840 attgaaaagc agttgtaatg aatcttaggg ttaagaaaga tcaagataca gaaatatcct 33900 tgaattgaag ttccaaaaca atgttgtttt ggtttttgtt tcattttgaa tcctttcata 33960 cttaggaata ccatttctag taaaataaat attttatgtt tagttagaaa tttatctgta 34020 tttcatacat acttagtact tttggactaa gaactgctat ttgaagtata tttgagggaa 34080 tgagtttgaa attttgggtg caggacatta taaagttgta actatgaata aatttcagtt 34140 atgcttatgc atagttttac ctagttttat ttgtctattt gagtattgtc cttgaattta 34200 aaattttttt cagccccaac tgatacacac acatatacat acataataca tgtgtgtgtg 34260 tgtagcttac agagtgttta taggaaactg attttgtata ctttggctac tttgttgtaa 34320 gttctagttt tttttctttt attattaaac tagtgcacga catcaatgct atatgattgg 34380 tgtttcgttg acctagaaat aatgcatgcc atcttctttt cacagctgtg tgccaaccac 34440 gatgcaaaca tggtgaatgt atcgggccaa acaagtgcaa gtgtcatcct ggttatgctg 34500 gaaaaacctg taatcaaggt aggaaaacag tctgacataa atacacaatc gaagacacct 34560 ctatcactcc caaattaaaa atattcttat ctcaaactac tttccatggc tatttttcca 34620 aaatatgtga gctgctattt tgctgnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 34680 nnnnntttta tcaaaggaca attaaacaaa ttgtatcctc ttattctctg atactaatta 34740 aaatgtattt tgaagaaaag aatccctgca tcagtaattt agaagccttc tggtactcct 34800 tgtttttcca ctaactagtt ggaaatcctt ggccaaataa ttaaccacct cagaccccag 34860 gtactgcttc cctttaatgc caaagtcaag taggggattt gatttgaaat ttggagtttc 34920 ctcctaactc tgagcccttc gattcgtgat taaatctccc ttcaactact gaccagtttg 34980 gaatgtttcc atgataaata aaaatgatta atttagcaag cactttttaa aaaaatcagc 35040 atcagttgtt taaagcaaat atttattcaa cactcatagt gccccaggta ttttatatat 35100 ataatccatt tttatcttta caacaacatt aggaagtaga gcattattta ttataatnnn 35160 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 35220 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 35280 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnntttt ggataaaaaa atatatttgg 35340 aaatgtgatc ctcgaactca tgctacaaag tcagacaagg cttgtcttgt taattaaatt 35400 cagttaaaaa tttccattat tnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 35460 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 35520 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 35580 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 35640 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 35700 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn ncattattag gaattaaact aaagaaagat 35760 tagggttaga tttcactaaa tagataagtg tcaaaaataa aaaagataag tcaattttta 35820 cttatttttt aaattatact ccctatcatc ttaaatgtca ggtgaaataa tcatggtgtc 35880 taactatccc ttacatactt acttgacctc atctgatata gaaatgatat tgctgaaata 35940 ctactgttct tcagtgtctg atacttattc caagatactc cttgaggtat gtcatgtaaa 36000 ctaatgattt atagacacag tttttttttc actatttatn nnnnnnnnnn nnnnnnnnnn 36060 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 36120 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 36180 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 36240 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 36300 nnnnnnnnnn nnnngacacc gtttttacaa tcatccagaa cattgatttt tgaaaacata 36360 ctcaaaatgg tggttcatga accatcagat gaaattattc atgaaccatc agatgaaatc 36420 cacactagca aaaatgacat gtcccaatgg tagaagctgc tattctaaag tgtgttcttg 36480 tattatctgc tatttgattt agaactgatt ttcccaatgc ttgttttctc tctaatactc 36540 tttttaactt gaaatttacc aaacatacct gcataatctt tttaaataag tgatgctctt 36600 attatctcag tttgctcctt aaaaaactcc acttgatttt tcttccccag cataagtttg 36660 caggtagcag tgttctggtt attggatgcc aatgttcatc ttagaatctc agtatacttt 36720 ttttaaagtg gtgaacataa gccctcagag gtttcattaa tatatcagca cggtaaaata 36780 ttgttgtcca aatgtgagga tataaaatat aaagaagacg attaaaataa gtcacttttg 36840 agattgcaga gaaattcatt taaatttctt tgcagaagct gcatcaactt taacatgctt 36900 taaagacatg ctaaaagtat tattgaaaca aaagttcatc actgggtagc tcctattttc 36960 agaaaaccag tttaacatgt actttttttt cctgtaacac atacctattt ctctaaagaa 37020 aaaatcgaat gtatcaagtt aagatcttgc tccccaaaac catattcctt aaagagaaga 37080 ggatttgatt agtagcaaaa tgggatttta cacctgcaaa aatagtgctg acactgaaaa 37140 tgtaatttca agtccagttt tgaaaataaa actacagctt aaaataaact taggtgtttt 37200 aatcatctct tattttgccc tcctcacaaa aaagcagtgt ggcaggttcc tgatggcaag 37260 gttnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 37320 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 37380 nagtttcatc taatggttct tagtcctgaa cactgtggaa gcttgaaagg gttccaccag 37440 ttcttggtac agcatcttag agagtctctg catccataga cttgagggcc ctgtcaggat 37500 cagggaagct gccatgtgtg gcctggttga gtgtgagagc tgcctagaga cttccataaa 37560 agttgttaga gaaaattgtt gatagtgcct acatagcaga ttaacttaaa ctgttttcat 37620 tcaggcccac acattttaat aaagtagaaa atatgcttca cagataaggg aaatcaaaca 37680 ggctcctttt ttctggagga gagaaatgtc aaaaagaatt aaatttgaaa taactttaca 37740 gaactggaaa ttagcttttg attaaaagta gcttttggta tatgacaggt attcactgag 37800 aattttgtag cgagttatat actttaagaa ataaccccca gaaacttgca tcatggtgta 37860 aacagcttga ataaacaagt gcttaaccag tgcctttaga gctgcctggg aaacagccag 37920 aataccaggg caagctgcat tttggaactg gtttaattta gtagccttgc cacaggctta 37980 gtgtgatctg cttttggtgg cttgatcttc cccactaagt cattttctgg atttgttaca 38040 cctagaactg ttaggaaatt acaggcttgg gctgatcatt aacatactgt actctacaag 38100 gcacacgtta cctttcaaag cagatgaaat tctaacctga attctggcaa gattctttga 38160 tcatttgctt cctttacttt tacttttatt tatgcatatt tccctcctcc ttgagtttct 38220 gtaccaacac aaacctcttt ttcccccagc cagnnnnnnn nnnnnnnnnn nnnnnnnnnn 38280 nnnnnnnnnn nttttccatt ggggaatgat ttttagtatg taaatatatc actgcatatt 38340 ctcagaaatg aaagacattc ttaggaattt acagtgtact ttataataat ttcagaagaa 38400 aatatttata aatgtcaatt cctaatgttt tagcatggtt tatgtttcat atgttgaatt 38460 ctttatcata aggaaagaat tggagtcttt taggtcagaa ccagatacta attttgtgga 38520 ctagttacat ctgaaagttg actgctttgc taagcacaaa aatctaaggg ctttaactct 38580 aatattaagg tggttaccta cagccgtagg ttttgaaaga tgtatggttc cnnnnnnnnn 38640 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 38700 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 38760 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 38820 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 38880 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnngaag 38940 ttgtatgctt tcaactccca aaaccaatgt tcatttgatt tggacattta actggctatt 39000 aataacataa tggtgttgaa acagaacgtg attgttaagt ctcaactctc ttctttgagt 39060 ctcacccatc ttatattttg gtaagctaat aatggcaaca gttaacattt ttttgcataa 39120 agcttctcat gtatgttcca gaagcataat ttcttacatt taaatgataa tattgacgat 39180 ctattatgct gttatttaaa taaaatcttt atgtaaatca tacatttgtg cataattttg 39240 gagtatttta aactactatt gtacaacgtg atagtgtagg gaaaaaaatc ttaagttggt 39300 cgtttacaat tcagtgtcct taaatagagc aggtggatta taatcaatat ttgtataatt 39360 tgttttttgt tttcttcctt ctgtgtatga ctaaataata cttttaaaaa tgatgatctc 39420 ttgtttggaa ttttttaaaa aattgtatga cgatggattg ttcatttata atgtgtgtgt 39480 tgtccttggg aaacaagatg acagtgatac atttttaaga aaattaaagg aaaagagaaa 39540 ccagaagaac cagaaactat tgtttataaa gtatctaatg cattcttttt aaactctacc 39600 aactgtatgt aattgtttaa tatatacttg caaagatttt tagggctaaa attgacatca 39660 gttcaaagtt gactcttatt accttcttcc tggtgtgaag acgagcacat cccagctcct 39720 cttgaccaag gcagtgaaca gcctcttttc caacccctgg atcaccaagc cacaagtttg 39780 ccttcaaggg gtgagcgtgc attgtcccag gtggtctgct cttcccacct gcatggcttc 39840 ctgcagttat gtcacttctt gcaaatgttc tgaagactag cccaggagtg tccagccttc 39900 agccctgagc atgagcaagc agcaggaatt gccacctggt gctgattctc tgtgctctga 39960 aaagcatgga gccctactgt ccttttaatg catcgagtat cattgctttt ctttctcttt 40020 tctgaatttt gaaatatgtt ctaaaaacag atagcaaaac aacttttttc atatgtaggc 40080 aaattcagga gttccaaagc aactgaaatt gggggtttga tgccttttgc caaggagatt 40140 ttagaaatca agtcctcttc cccactcttg aggtatagag ttttaggtta tctgaaaatc 40200 aaagcataca ttagtgacct tcttttggta tacattgtgt aacatcagat agcctaagtt 40260 ctggttccaa gagagcactt agttgctatt tgattataag caagtcatta accaatcagt 40320 aacagggaga cagtaattcc tgccctattt gcctgtgggg ttggcgtgag aatagaaaca 40380 ccaaaaatat taatgtcttc gtcatgctgg aaagctttgt aggaaaataa tgtggtagca 40440 taattttaca gttcctcttt taggtcattt tatctctaac ccattcatag gtttaagaac 40500 ttagaataac tgaattaaaa tagttgaaat tattaatcat tgtctctgct gagaagaaat 40560 ttgatgcatt tgtatatttt ctacacaagg aattagggca aaagaattaa tttcggtccn 40620 nnnnnnnnnn nnnnnnnnnn nnnnngtagt tgacatattg acatctgttt gcctatgaca 40680 ttgcccgatt taagcaccaa agcggggaga aagtccaaat gtgttaaaac aaatgaaagt 40740 ttatttttaa aaaatccagt aagttagtta tctcccgatt tttcaagcta cttttcagtg 40800 tctgcatgct aataaaattt ctgatttttt tcctgaagtt taataataat gctgtgcagc 40860 ttgcatttcc attctggacc agttcccttc tccctctcct tctcccatcc aagctcctga 40920 atccaccaac ctactgaaat gtattcctga caatagtaga tgcactttaa gacttgtata 40980 cataataact gaagcatttg aatgtaagtg gtttattgta aagttctatc ctttagtgta 41040 agataagcac atgaaataat tcattaaatt ttttattttc ctattttata gattttcctt 41100 atatatatta taaacctcca gagaaaaagg aagataagta aatttaaaat aaaaacacca 41160 aagttttatt tctaggttct tttatcaact tttaagattt atttgagaca gtatgatcaa 41220 tgacttcatt ttgttctgct tattattgta ggagtattta ctataatttg gaagtaattt 41280 atttttgaat ttattgctta attgaatgat ctccaataga ttgtgataat gaacacagca 41340 tttatagaaa gcagcacata ttaacttact taatatggca ctaggtcaat gagaaaagaa 41400 ggtaacataa ttgaagacaa gaaactctta agaaaactga ggacaaaaag gcttctcacc 41460 aggacaccag atgcatttaa tcttttgaag ctctgtactt taggaaaagt ctgatatttg 41520 gcaaattttg ataaacatgg atgactatgg aatcctattt tatagtatct gaagtggctt 41580 tcataagggt cattgtgaag tttttaggag acacctgcct gtggcagatg ggacaatgat 41640 ggcagtcact agtgatatta acaccagtca gctgtcaggg aatatcatcc agaccatcag 41700 cagctggtag agtacagctt tctcaattgc tttccatgtt ttggatactt atatgcccgt 41760 taataacagg taaaatagcc agtacatcat ttccacattt acccattgaa tgttgcatgt 41820 tttcttcctt tcacatattc atacagtcca gatttttttt tggactcatg acagcacatt 41880 ggcttttctt tcctttcagt ttcatgattc ttaaccccaa agtgcttttg ccatgggaac 41940 ggaaggataa atttcggtcg aagccattct ctaaaaccac cagcagctcc agtccagatt 42000 atgaatacta ccatagcaat cctaaggggc cctttattgg cccagactgg agccaaattg 42060 tagaagagct gccccaactg ggcttcaggt ttagttcagg gacatagtgt ctgaagaaat 42120 ttccatcagc ataaatactc cttgtttatg agctgcctga aactgtaaac accgaatcca 42180 ttcccatcag gaactcacaa aagtttctgt tatgctttga aaataaacct agggatactt 42240 aactgacatg taaagaaaaa tccatcagta tccttttctc gagatatagg ttttgatttt 42300 actgtgttat gttgtggttt tgttctgttt ttgtttagtg cagaaaaata cttaaaacac 42360 aaagcctttc tctaacacca tatttggttt taaatgctat ttgctataat atcaagaaga 42420 tttatcaaag acaggtgtga ctctgcagga ccatattagg aacagtcatt aactcctctt 42480 agaagatgag aaaagtttcc ctctcatttt tagtaaattc tgaaatacaa agtagaagaa 42540 tatgggaagg gtagaaaatt tctcagtcat tcttccttga tttttgcttc aattaacaga 42600 gggagaaaac atttttgata ctgtaatctg catggcgctt tctgtgagga aatttttttt 42660 gaaatattta aaatgtgatt ttgttttaaa aacccttaac tagtcaagag acaataaagt 42720 agaaaattga agaacgcata ggagatgaca aaagtatttg gtttatttat ttaacctttc 42780 cgggatttgc ccagctcggg gtctgcttac agttgcattc agggcagata aaagacctac 42840 ttggaaaatc agtaagatat gaaaattttt ttagatgtaa taattatttg gtggttttaa 42900 aacataatgg aacttgatga tttagttaaa tgaaatatac agatttctaa taaatgagca 42960 gatcagattg aaatagatta gatttgaaaa caatttttgt cactagacat attatctata 43020 ttttttattc aatgacatgg attaatagat aattaaatta tgctaaatgg gtacttatgc 43080 tggtggttcc agacaaacat gtggtcacat ttctttttaa tgaagcatat attttgggca 43140 ttactatttc aatgtttatt tgtcattatg agttttatgt tcacttgagg cagtgtttaa 43200 tttaggttaa aaccttttca taatgtaaaa tttgttgatt catttcatat tgatacttaa 43260 accaataaac ctgaaataca tgacagtcta ttacattttg tccattagtg tttccatatg 43320 ccttttttaa ctgcggaata aatgaaaata tgttggtagg ttttttaaag ttcattttgt 43380 gacttgaatg cattactaaa gtaggaaact gaagtttctt ctttaatctg aaatcatata 43440 aaatctacca gattgaacaa gaaaaactac aatattgatc actcttaatt tatttcgttc 43500 ttatcctctg aaaatatagc ctatgtagcc ttcctttggg atgggaaggt caaagcagtc 43560 aaatgtttta aaatctgtat acttcctgta aaataaaata ttctcaaagt ccaagcctta 43620 aggaggccaa tgccttatcc atttaaagta aaatattctc atgtaatatg ttctttaaac 43680 agagaaagga agacattgaa cgaatagggt tacattgtca ggattggatc tagcaataga 43740 acccaaatat tttgagaata ttggcaaaca gttgttagct gatagagcat ctctgcccat 43800 gaggacgtta gtatgctgct gtttctaggg ttgataaggt aggcacctgt tgttgagaat 43860 tttcctaact ggcctttata aacagagatc cataaaggtc atggcgactc ctggttgccc 43920 ctagtgtaac gtaccttcaa attaatattg catgaacagg atctatctct tatgtaaatg 43980 acagttactg tagcttgacc tatttttctc tttcattttt taatatagaa aagaatttta 44040 gatggcttac attagtgctg aggttttgta atttttctgc ttcaacccac attgttttca 44100 gatctaaatg agtgtggcct gaagccccgg ccctgtaagc acaggtgcat gaacacttac 44160 ggcagctaca agtgctactg tctcaacgga tatatgctca tgccggatgg ttcctgctca 44220 agtatgtcaa gaatcttaac tgttttataa gtgctttggg cttgtttctg ttgtgctctg 44280 agagcttgct tttgtgaaaa tggcctccgg ggttctccta aacaagatgt gtgcgtgtgt 44340 ttatactttt gcctgaggaa ttgaaaatca aaataagagg caccattttc gatgtataat 44400 actcttctat cacgggtgcc aatattaaat tgattaggaa atggtgttta gaataagaat 44460 tacaaaacta agtatgtttt ctaatttttt ttttaaacat gtttacccat gtcttctatc 44520 tgggcataga agaatgattc caggctaaaa aaaaaaaaaa aagaacacta ctgactattt 44580 agggactgct cagacagaat ccatcaggtc tgttttgtaa ttttaaatca ttcataaata 44640 ttttcttcat ttttatattt cattataaaa gcctttaggc ttttatagaa ttttagactc 44700 taataatagt ctaagacttc taaaaaacaa tttcaaataa aaaataaatg aatatgatac 44760 cagaaaagta ggattcatgt ttataaggaa gaaatgcatg ttttcaccat catttggatg 44820 taaaaaatgg actttgccct aaaattctct ataggctgac tcctgaattg tggtattcat 44880 atgatatctg acaattaatg acttgatttt tatttttact gaattatcat ttaactgagt 44940 tgcataaaat aaagttagct aattttattt aacattgcat aagtatagca tggaactttt 45000 tggaattagg tgaatacatg tttaacattg tgcaactcaa tgggaaatct gctgttcccc 45060 tagagaaatt tcatgggcat ttgagacagt tacttggatg attagttaag aatagcgttt 45120 agcttgagta atctggaaaa atacctcaat tcttcacttt ccttggccac tggaaaaatt 45180 tccagataat ctatatatga tagttttata tttgtctttt aatgctttct ttccttcccc 45240 cttttatttt ctttagatgt ttgatagtta aggcctcttt gtcttcattg ttgttcagtg 45300 tcactgattt caggacaagt gaaaaaacat agctatttcc attacagatt ttgttttcta 45360 cctgtctgag tcagccagtc actctctttc caggtgccct gacctgctcc atggcaaact 45420 gtcagtatgg ctgtgatgtt gttaaaggac aaatacggtg ccagtgccca tcccctggcc 45480 tgcagctggc tcctgatggg aggacctgtg taggtgagtt gtaaaatcaa gcatctctgt 45540 cagcagcctc tgtaggataa agggagaaag tgaaaggtga tgggaataag gaaaaaaaag 45600 gcaattactt acatcagata attagctatc gttcagaaga tatcagatgt ctcagaagag 45660 gacatctctg tgaatggata atgggagcgt tgttttttaa aaaatgagaa atannnnnnn 45720 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 45780 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 45840 nnnnnnnnnn nnnnnnnnat tgtatgtgtt gttcttcagg cctcagattt ttttctggta 45900 aattaggaag gcagtgtagg attgagagtc tggannnnnn nnnnnnnnnn nnnnnnnnnn 45960 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 46020 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 46080 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnna agtgtttggt 46140 gattagagtn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 46200 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 46260 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 46320 nnnnnnnnnn nnnnnnnnnn nnngtctatt agccttaagg taagacctgt atcattagcc 46380 atgtgaagaa acagccaagc agctgaagac attgcagggc actgcggtga gagagaggtt 46440 ggcctgtgtg aggtttttca aggaagaagg tccatgtagc tcaagtacag ggtgtgagta 46500 tgaggggtgg tggctggaga tataatgggt gaagtaggtg agggtgattt gttgcagggc 46560 cttcaaggac acaggggagg aatgtggatt taattcaaag tgcaagtaga aatagttaaa 46620 agattttgag cacagtaatg atctgattgt atgttttaat ggaatcattt tgacagttgg 46680 gttggatatg ctttggaggg ggtaagtaac agcaaaagga actgttttga ggcttttgta 46740 acagtctaac ccaagagatg gtggcttcaa atagggtagt ggtggtggat taagagaaaa 46800 gtagacagat tcaaatactt ttttttagag gtagaattaa ctggacttga tgatatggga 46860 gttaaaggaa agagggttgt ttctaattca agaatgactc atgtttctag tttaagcaac 46920 ttggtagata caggtgcagt atgtgctaat atatagagtg caagaaaata ataacttggc 46980 ttagaaggct gggttgaaat ttggatatgt ggaatttatg ttgcctgtag ggacaaccag 47040 ctagataccg tttatgatga gccactatat gctaggcaac catttgtagt cagttattta 47100 ttttgaaacc cactctgtct taacataccc ttgctcttcc taaaatgcta ttgacttatg 47160 tttctagatg ttgatgaatg tgctacagga agagcctcct gccctagatt taggcaatgt 47220 gtcaacactt ttgggagcta catctgcaag tgtcataaag gcttcgatct catgtatatt 47280 ggaggcaaat atcaatgtca tggtaatgaa acccaaccat tgctttgtgt tgtttcttcc 47340 tagagcactg aaaggtctcg taattgtggt gatggctgga atgtcagggg caggggagag 47400 tactggcgtt aagttaaacc aacagacatc cagtttaacc actggtagtt ctcagtctac 47460 atgtagttta tttcttctgt ttatctgcca attttatgta gatcatcaca ttgccaaaaa 47520 aaatcatttt tgaaactgta tatatttttt atgtcatcat atttatctcc taaataagtc 47580 tcttcttttc ctactttctg atgcagacat agacgaatgc tcacttggtc agtatcagtg 47640 cagcagcttt gctcgatgtt ataacgtacg tgggtcctac aagtgcaaat gtaaagaagg 47700 ataccagggt gatggactga cttgtgtgtg tgagtagcac ttgtctctca gctttaaatt 47760 ctagcaggaa atacaggatt acacaaaggc cattgctagg gaaaataagg aataagatta 47820 tcaaagaagt ataattgtca taattggtta tatttgtctt tgatttccac aaacaataaa 47880 atcacttgct caggtacttg taaaaactaa ggactcagta atacactata atcttaagag 47940 tattttaatc tcttcactga aatctctcaa tattttcttt ttagctaaaa agaaattatt 48000 gagtcctccc tgagaatttg cttttgttaa aattagaaat catgtttgcc attagcgtta 48060 gattttgatg gtgggataat ctggatatat tctacatttt tttccctctg ttttatgcct 48120 cctaactctg ctattaaaaa atcattcccn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 48180 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 48240 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 48300 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 48360 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 48420 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnccat tcccatattt atttcctcta 48480 cattcagaac tgttttgctt ttaaaatttt tataagtttt atttggagta gtggttatgt 48540 aatagataaa catgaaaaaa acccaaaagt cataatttaa catacatttc atgtaacaga 48600 agataatttt ttaaatttcc tccgtaaaat atttactgaa aaccatggtg cataaaatca 48660 tgaggattaa agggttttta aagagtcatg tagtcattcc cagtctggaa tgtatcattc 48720 aggcagatta aatattgatt ctttttgatg aagattatca aggaaaaaga tttcacagat 48780 gatctttgca acctctgaaa atgtctattt aaaagcagtt gtatctatca gtaaagggaa 48840 ataaaaaccg accataaaac aatatgtaga acatttactc aagctgtatt tatgcaaatt 48900 gaatttatat aaagtgtgga taagaaaagt atttgtcttt agacaattct gaacacaatt 48960 ttataatata cacgtagcat agggatacaa cgtagattct aactatgcat ttgcttatat 49020 taaaaaaact tagannnnnn nnnnnnnnnn nnnntctgtc agtgtaatgc ttcacttttt 49080 agggaggaat ggttacatac tgatttaagt ctttgactct ttcagtatat cccaaaagtt 49140 atgattgaac cttcaggtcc aattcatgta ccaaagggaa atggtaccat tttaaagggt 49200 gacacaggaa ataataattg gattcctgat gttggaagta cttggtggcc tccgaagaca 49260 ccatatattc ctcctatcat taccaacagg cctacttcta agccaacaac aagacctaca 49320 ccaaagccaa caccaattcc tactccacca ccaccaccac ccctgccaac agagctcaga 49380 acacctctac cacctacaac cccagaaagg ccaaccaccg gactgacaac tatagcacca 49440 gctgccagta cacctccagg agggattaca gttgacaaca gggtacagac agaccctcag 49500 aaacccagag gagatgtgtt cagtaagtct aataaatgtt agcacatttt caataggctc 49560 tttataatga cttttcaacc acaggccatg ccttgaataa gaatgaaact cgtaagaaga 49620 actagctatg taaagtcgta tgtccctatt gacaaatatt ataaagagct acataaagag 49680 tcagtctaat tgggcaagta agaaagaatg tatgtagcaa tggaaggaat attcacaaag 49740 tcatatggta gatgacagct cttagccagt atgggaattc tgacatagtt ggatttactt 49800 gaaaactctc agaggtggga acttaactgt attccatgct atgttacttt taatctaacc 49860 cttcnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 49920 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnngg tgggaaagat 49980 ggttactcag aagataccag tggtagtatg ttagtgaaag agatctgtgt aggacactat 50040 gagaggagaa tgaagggata cctcgtcagt ctttgggtag ggaatattca gggagggctt 50100 cccgagtgaa acggagcaaa gtggatgagg aaaggaagat attatagtta acagggacag 50160 cccaacaaat gcaaagaagt aggaagcagt gtaataggca gtagactgcc agtaaattgg 50220 agggtcatgt cagggacaga tgagagaagc tggacagcta ccccttggat tgctgtaaac 50280 tgatagttag acctacactg gggctggatt agatcaaaga ctcccagatt cttagcaggt 50340 gactaggtaa agggtagcac caccaatgga gatagaggat tcaggaagag aagcaagttc 50400 tgaaggagga aaaggttgtt aatttattac cagaccaaaa tttgcatgtt ataactttca 50460 tctgttcctc atatttatct gcttgtcctg tgtcacttcg cttattctta attacctttg 50520 cattcactct tcctgtctcc tcctttatac attaccagtt tgttttgttc cttttctcat 50580 tcctctatga tgaagctaaa gtttgtgcct tttatattga atacctttat aactttttat 50640 caggtgaaaa aaatactgac atttattatt ctttaaaggg gaaaannnnn nnnnnnnnnn 50700 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 50760 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnta 50820 ataaagggga tatcagagcc gcattttgct tgtatgtgac tctcaattaa attttctcct 50880 ctgggtaatt gtccatttaa agaaagaggg aaattctcta attctgatat agtcatgtat 50940 gttttggata atcaatataa tttttagggg aattagatgg tagctctcaa aatatacatt 51000 ttatatgtgc ataactattt tctgaaattt atgtttttgt aattttctcc tcaccccctc 51060 tctccatctc agatactgtc tttttctccc tattactctc ctttttaaat tctcatcatt 51120 gtgatcatac aatgggattt ttaatttatg agtgcttaag taattatggt atttacatta 51180 ttttgctgcc ttaggataac agggaaattt ggctatttaa tgtaagatga tacgctaaat 51240 atttttttca ttatgatgaa agaatacatc tttctgagaa ttttaaaaaa tcctttctct 51300 ttttaaaatg tttctcttta tttccagtat tcctctctat gccaatacat atatgaatta 51360 aaaatgacat tgaagttcac caataaaatt tagtgcaata aaattgggga aatacagagt 51420 tccaatgatg tttgggagca ttcattataa gagagtgtga tcttaaagac atgctctggg 51480 aaagcattgc cttgatcaaa tgccaaggct gttgcatgcc acagatagca ttgctgcctt 51540 tagaaactcc tgccaaaaat taaattccac tctcatttca tcttcagcga tagctgctta 51600 ttaagtctag catgtgtgag aatgctttag atgctttttg acttgctgtc ttgtggttat 51660 agcatatatt ctaaataagg caaaggctct aagtttttaa ttcaggacaa ataaccagtg 51720 tctattcact gtgaaaacag taaggaacct tagcactatt tcagttcaac accatttctt 51780 tactgaatat tttcttcact tctggcataa cttcttaaat catcctcatt ctccgagaca 51840 ggtctgagaa tagaatttag ctcacctcat ttcataaatt tttaggtgat gttaattatg 51900 gccactattt ggcagtctcc ttcaaatagc cctccacttt gtgtttttct ttacattgaa 51960 agatattttt aaattgannn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnng 52020 aaagatttct tttaatggaa gaggttgtga aagagagctt ttccttataa tgtatgtgcg 52080 ttgttgccct gagaaaagac tgtaaagata ttctaaaaga aaatcaagga agaaaaatat 52140 tataacaaga acacatcttc agcccagacc tctcccccaa actctagacc tggatgtcag 52200 ctgactgctt aacctatcca gttggatgaa aatagacgtc tcaaactcaa catgtataga 52260 attgttcctt ttcttccctt aaacatcctc tactctcagc ctttcttatc ccaggtaatg 52320 ggacctctat cacctgttgc taaggtctaa taatgtggat tcatccttaa ttactcttca 52380 tacatacagt caatatgtaa ggaaatcctg ttagctgtac cttcaaatta tatttaagtg 52440 tgaccttttc tcaccaactc cagtgctacc accctggccc aagccatctc tcccctggaa 52500 tacaggccca aaatccttta tccaaataac ttatggaata tagcattttt tagattttag 52560 aaaatcagta aggtacacat actatatagt acacactgaa gtagtgaaac atgctgattt 52620 tcctctagtg cttttactgt gaacgtatca atgttaagga aaggaaaatg atattaactc 52680 agagatgatg tctcacagca tatatttact agcttgcaca aattttttaa tgttagcaag 52740 atttaaggca aattttttac ttatatttta attggatcct atgatgatta ttaaagaaaa 52800 aagtagttat ctctagaaag tataaattga gctcttggca aatgtgagca aaaccagaaa 52860 tcagattttt ttaaagttac gtgtacattt gtctatagag ttataattaa aagtattgtg 52920 ctcatagcca gtctgtgatc tggggcatat tattatatct tcctatgcct taaaacaatg 52980 tttcctgtaa gtataaacaa agtaccattg acactgcagt ttttgtccat ttgctacttc 53040 caaaagaaag gggtacaaaa cagaaaagtt atttaaactt taagcagttg gttagatatg 53100 gtagttataa tgagctttgg tattttaata tggggcttaa aaattttcat ctaaaccaac 53160 attaattatc taagtgtgat atccttagga ggtctttgtt aagtcctccc aattatatcc 53220 agcatctcca aaggtgactg aagtcccttc taataactca tcnnnnnnnn nnnnnnnnnn 53280 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 53340 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 53400 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 53460 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 53520 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 53580 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 53640 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 53700 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 53760 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 53820 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 53880 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 53940 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 54000 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 54060 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 54120 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 54180 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 54240 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 54300 nnnnnnnnnn nnnnnnnnnn nntcatccca ccactggagc atttttcctt atatttaacc 54360 taaatttatg aaactacagt ttatgcattt ttaatattgt cagatactca gggttaatgg 54420 acacagttgt tcattgtctc cagatcccat actccatgta ctcaaagaca ctttcaggag 54480 atgactcact cttccctggg cccnnnnnnn nnnnnnnnnn nnnnnttttc ttagaaatgc 54540 tttttttcaa cccacttagc aatcttgttg actggtcctt tgaacattct ttattttcca 54600 aatctgtcac aaatgatcga gtcaaagata tataatagtc aaatctgacc caatactgat 54660 cacggggggg tttcttttaa catgttttgt aatctaaatc atttctcttt gttgcaaagt 54720 agagatttat gggcagctat ttaatctcca gatgtttttt cctcaaannn nnnnnnnnnn 54780 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnntggaatt ttatattgga tttaattttt 54840 ttcattctaa gtaatccttc atatgcttaa ctagctaaat tttacttgtt ggttctgata 54900 ttccttcccc aaataatcaa agttaattca gattatggtt ttttttctta tcctgataca 54960 gcttaatata gccattgaaa atattctcta gaaaattcta agtctaatat acttctaaat 55020 tttaagcatg acactgaaaa atagtgaaaa gcatagaaaa ggaatatgaa aatagcattg 55080 aaacatttgg aattaagggg ttttaaccac tactaaagat agaatatact gaaccagaaa 55140 acctagtctt gatttctacc tctatttatg aggtacatga ctttggacta gtatgagtnn 55200 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 55260 nnnnnnnnnn nnnnnntccc agctatcctt tctcactgag tagcttcaaa gatcagagat 55320 aagggatcta gtgtttgtga attctgagtt actatgaaat gtaaaagatt aatgtgtttt 55380 gtttatcact gtaaataacc ttgatgggga gaacttatgt gatttaaggt tgcaactttt 55440 tttgtgtgat gtgaaatatt aaacatattt ttaaaataan nnnnnnnnnn nnnnnnnnnn 55500 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 55560 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 55620 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 55680 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn aaaaacaatt ttagcttgag 55740 ggaccctagg tttttttagg ttatatgact agaaatataa taagatgtat ttacagggct 55800 aatccggtgg gcatgatttg agcaacacta aagttatctg atgtagcgta aaattcttgt 55860 tgtttcatta atgagacaga cttgtgcttc tcaaagtgtg ttttatagag cattatttct 55920 ctgaaatatt acacagttgt atgtgttaaa aagttcttag gtcaaataca tttaagaact 55980 cctaggttta gctaagttaa ataggtttct taactgtagg acttctcaat actccaatgt 56040 gctaacaata ttgcaaatct ctaagagagg catataatgt atcttttccc aagttgtttt 56100 aaccatggga ttatttgaat cttgggatta gagtttactg gaatcaactt taggtcaggc 56160 tgggctaaac taggaagtat ggagaccata ctccacaaaa cccagtatcc ctcactagtt 56220 agctctcgtt gcctggtttt tgagactaat gggttgatct tagatttagg aatgagggtt 56280 ggggagggca gaaaggtgct gcacaaactt agccagaaaa gcagccataa gtggtggtat 56340 gcctgtatgg acaccatagg tttgctgcag gggattgtta aaaagccact tttcaaagac 56400 aaaactctgg tgtcacaggg gtaggagcaa tcccagggca aagaaactca gaaaaacttt 56460 ctgcagaacc caaagtaaag gaagttaggg ggacatgctt tctgttctag caatatataa 56520 tgaaatctta tataactatt aaacatgttt tatggaaaat atttaataat tttggaaaat 56580 acccatgtta caaaaatgtt acctgaaaag tagaaagctg tacctgccat ttgactccaa 56640 ttagatataa aatacatgtc tgcatatgtc tggatgtgtg taaaagtctg aaagaaaaaa 56700 gaccaaaata gtaaccatga gtatatcctg tttatgatat tttgggcgat tttcattgtc 56760 tttttgttgt ctttatccag taatttttaa actgttcaac tgtactatcg ttgataacag 56820 taattcatta tatcattaat tccttcagag aaggggcaga gtaacagaac cccactcttc 56880 tcagcaggaa agtgaggatt ctgattctta atcatgttta ttttattttg gacatactga 56940 gttttaaaaa ttcattgaag gttttattaa cttctatttt taacctttca gattttcctt 57000 tacaattgaa tgtgatagat atgtccaatt ttagctagga aatttgtatt acgagaggga 57060 atataacatc acttggacca ggaaacagat tttttttccc cagtgttgtc agaagcccat 57120 tttaactaaa ataacatagg atgtttatta actaaaataa cataggatga ttatcttaag 57180 atgtagattc ctaatcctta tgacacattt actaaatcag acttactagg gaaagtatct 57240 ctcagtcttt atcnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 57300 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 57360 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 57420 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnntctg agtaggcaca cttagtcccc 57480 aacactcaac cttttattca accagcggtg agctggatgt gaacatgaca gacccagtag 57540 ggttccaatg cctgacaacc tgcacctgtc aggaagagcc ccctcctttc ctgctcccct 57600 gcaacacatg gttannnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 57660 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 57720 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 57780 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 57840 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn atagcagggc tctttctaaa 57900 gcataatggc agctccaacc cagcagaacc atattctgga tggagtcaga actagccaaa 57960 tgtaggctat ttaaaatata gcaattgtgc ctaggcatgt gtccacctga ctcatgtaaa 58020 ataaaaagaa aaggcgttaa tagaagccag attatagaac ataatgttat taatatgtat 58080 caggtgctct tatgagtggc catatcagag aaccagcctc attgttgctg ttattacgca 58140 acatgagact gtgctgcaat tccaagtacc aagcagtaag aagaaaattg ttttctttta 58200 ctgattgctg catgttgtcc tgaggttttt cccctcatct ctcttctaca gattttacag 58260 agtctgtggc tagttcagtc acattgttct tagtcatgga aatattatgg tctttctgat 58320 tctgtgtagt gataagtaaa aagattgttt ctgctgaggg gtgaaaaggt cttcaaagta 58380 gtttgctttc ttgtaaacag taagacctgc agagaccctg agaagatgcc tgatatctcc 58440 ttgaaaatta aatttctgct agtgttttga aggagcgaat tgtcacttct cacaggttag 58500 gatctgctgc tgtgaattct gaaagttttc aagatttttg atttatattt taattagatc 58560 ctatggtagc tagtaaagaa aatatcattc tctccaaaac gtataaatgg gcccttggtg 58620 attgtgacca aagtcagaag tcagattttt ttttgaagtt acatgcacat ttgtcaatag 58680 agttataatt tacaagtatt gtcctcannn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 58740 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 58800 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 58860 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnctac tattgatgtt gaattcattt 58920 atatagtaat gataacattt cctacttaat tcataaaaag acagcctatg ctgttttctt 58980 gttctgagtt tatatgtttc tcatgctttt tattatggtt cattacaatt ttaatgttat 59040 ttttaactaa ctagatcctt ttgaaacaaa ttggtttgca agtgtgagct gttaggtgca 59100 cagagaaaaa tgaaaataga aacttgcgat tttattctag gcttgttacc aaatatttag 59160 aatactgtgt tttatttagg tgtttatagt ctcattagac agttgtgatt ttaaaataga 59220 gaccacatca tctcaacttc tttactgtga aaataatgac aatagtcttt tcagagatga 59280 atctgtctag atgggaaatt tacatgattg atctgatgag nnnnnnnnnn nnnnnnnnnn 59340 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnaaa ataatctgac aagtagtttc 59400 cccagaaaat ctgatttagt aaatgtacca aaaggattta gaaatctaca tcataaataa 59460 acattctatg ttattttagt tcagacccta ttttaattca gacttcctat gggataaaaa 59520 cttccattct ttctttaaat agattctttt ggcttgagtg catttacacc tgtccccaac 59580 agctggtggg cttctgctca ccctagacgg tgttcatgct gcactcagtc caagcagccc 59640 ttatcagaga gtcttcttac cacttgcatt ctggtgcgaa ggactcattc ctggcagagc 59700 ctaaccttca tgggaaccat tggcttagaa gagaggagaa gnnnnnnnnn nnnnnnnnnn 59760 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 59820 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 59880 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 59940 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnntatcc 60000 tgtctccggg cagtgtctct gagtctttct aaaactttat taaaagttcc tacaatacat 60060 aaaagagaaa taggtatgtc acaaaaattg ggctcattgt tgattagcaa tgtctatgct 60120 ccacctttca ccagagattt aacttttttg ccaattttgc tctttactgt cagcccaaag 60180 gtgtctgtgg aactttgtag attcttccta tgggatgaaa gctttgacaa acaaggtctt 60240 atttctgtga gtccaacaac ccttttcttt tggcttataa tgaatagatg gctaaaactt 60300 ttctctaaca tggttaaaga aatttcagaa actaatttcc agccctttta tttgcttttc 60360 tctctgatca cttaaaattg tgatgcatat gtcctgcatc tgtnnnnnnn nnnnnnnnnn 60420 nnnnnnnnnn nnnnnnnnnn nnnnnncatc tcataaaact cttgtcattc atccctacgc 60480 tccaatctgt gtctctaaaa gcctctaggt tttgccacca gaaacagcct tcagaatatt 60540 ggaataatta catatgtacc attctcttca atgaactaaa ctcctaagca tagaagtaat 60600 ttaagacagt gttattttta atcatttagt ccaacaacaa atgccagaac tggcctgggt 60660 aatgccatag gaaccacctt gtttcctatg aaacaggaag agaatttgca aacctacctt 60720 tactaaataa tgcttacatt ttgctatagt tactcttggc aagaagttga gcagtgggag 60780 gtgtatatga taatatttac atttacttct ctgtgcttac tgttaatgtt ttctgggtaa 60840 aaatatgcaa ttgactattt gggaaacttc atttgtgaac acaggttata tatgatcaca 60900 tccctgggga aaatacatat ttagtagaaa gtgcctggcc agcatttctt acaagaactt 60960 tcctgctcct ttttgtggct gattttaccc ctgactccag ggcccagtag ccattagaaa 61020 gtactgtgct cctcagccca gtccaaccat gcctttatcc agctgtcact tgcttcagta 61080 cctgtacatt ccctttcttt tagtttatgc agaaggctgt aagaagcaac aggcaagaca 61140 tatcttttgg tgggtaaaac atggaccaat ggtataaaga ttctggggac attttctaaa 61200 aatatatgaa cattcatggt tgatgttaat tttagtcata attcccctat gacacccact 61260 ctcaccttct tcctgttcat tctccaagct actaccagta atggtcattt cagcatgcaa 61320 acctgatctt ggcaatcctc tgttaatcct tcagaggccg ccagtttcca ccagggataa 61380 cgtccaaaat atttacaagg ctttaaaagc tcctgcacac tctgcttcta tctccaattc 61440 catttgatgc ccttcctcct gcaccttctt gctgcctccc tccccctgta aggtctacag 61500 caattctgat gtttcagtcc cctctctttt cattctccag agccatagct catagtactt 61560 cttcttatgt catagagaaa gcatcacttc tcagagattt ttatgacgtg tatcctcccc 61620 agcctagctt agccagtccc tcactcccca ctcccgtact cactagcatc ctctgcttct 61680 ttaaagcaca cattacctgg ggaaattgtt aaaaatcnnn nnnnnnnnnn nnnnnnnnnn 61740 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 61800 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnaatgagt ggatggagta gaaggaggca 61860 ggatgcatgg acagatggtg gaggaattgt attgctagca cttctctgat tcttttttag 61920 tttctaaatt tataccattc taattctaag aattaatggt tcaactgcag agaatagcag 61980 gtatgcattt gtttttatga aatatatata ttgtgtattt aagttgaata ttagcccaaa 62040 gtatcaagca ggggaaagaa caattagaaa aatcatgtac caatgtctat tcctgggnnn 62100 nnnnnnnnnn nnnnnnnttt aattaacatt ttctgcattg atcaaaatag ctcccttgaa 62160 atcaaagaaa agtgtttgaa tttcacaaac atatttcaat ttcatagcct tgcgttcaac 62220 aagtatcaag cctattctaa gttctctttg agtaaccaaa atacaaataa cagacatata 62280 attgctattt atatgtgatc taacagaacc ctcttttatt tagatgtctt aggtaagttt 62340 tttattcata tttatatttt ttctttttac aatcctaaaa acatagaatt acaaaaaaag 62400 tgaaccaata caaatgaaaa aaaaaatttc atttctaata cttagattac caatatttag 62460 catattggca tattggtgta tttatccctc tttagtcctt tttttttggt ttgtgtttct 62520 tgtacatgat tgtgataata acataatact gcttgaatga tttgttctgc tacttatttc 62580 acttaatatt ttcacataaa ctctaaagtg tatcgggggt gggtagtttt ttttctcttg 62640 aaacttcagt gggatgctct tagtaatccc atactggtat gtgtgaggaa ggaaaattag 62700 ttaaataatt tggtatggtt atagagtaga gcacacaaaa ttgtaagaac caatagcttc 62760 tgagcacttc tggtctctaa attccttgga atgtttccca gtggattgta atgaaggtat 62820 acatgatcat ctgctgctaa attaaatggt tcttagaaac caagaaccnn nnnnnnnnnn 62880 nnnnnnnnnn nnnnnnnnnn nnncatctga cattttacaa gttgttactt gactaattct 62940 ttgtgttgcc atcttctaac tnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 63000 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 63060 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 63120 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 63180 nnnnnnnnnn nnnnnnnnnn nnnnnnnaca ggctcccggt gccaggaaac ctttacatca 63240 acactcgatt tgccatttga tagtccttca tctgggagga aaaaaaaaga cggaggggag 63300 cttgaaaaac tgtcataatg tccctggaat atggtacttt taagagttga gcctattcca 63360 ttttggagat gatttatata agttacaaca aaagaagggg acaaaaacat gattgttcta 63420 tggagttttt ataactttct gtcacaagaa agcacgcttg tctacaattt tgtaatattt 63480 ctagtaaata aaagaggcac tccccgtctc agagcaccaa ataaggaaag tgtaattgga 63540 tgtcattgct gtcagtcagc tgggctataa aagagagagt ggggttgcct catcccctgg 63600 gtatccacag tcagctgtgt ccctagagct tcttttcttt cattgctgcc cagctgggta 63660 tattgcaagt atggattata agaggggaag ggacttcact gttttaacgt ttgaaacaaa 63720 aaggaaaaaa ctcagaagta gtaagctaaa aacaacttgt gcaaacgttc tgggattatt 63780 acttaatttt aagaattttt gctaaaaaca ataggaacat cgttgaaaaa agaacccctt 63840 tgagtgatga ctgtttcata gtgttctaca tctcctactc cctgccttat aaaaataaag 63900 ctaattaatt gaagttctgg caaagagaag ggagtattcc ttggcctttg accannnnnn 63960 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 64020 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 64080 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 64140 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 64200 nnnnnnnnnn nnnnnnnnnn nnaatgattt aaagaaatca attatttttt taaagtagat 64260 tcttatgctt tgtcccactt gtgtcctttg aggaaaagtg tgaaatacta cctggggttc 64320 taacaggatc tttgnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 64380 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 64440 nnnnnnnnna cacatgtgtg taaatagctg caattcttta tgaaagtgtg taataggggc 64500 ttataagcat gaactcttga ggtacaagag ggaagaattt gattctgcct ggggcagaat 64560 cactccacag aggccgtgat atttatgctg ggccttgaag gaagaagaga tttttggcag 64620 gttaattagc aggaaagaga gcattgggtc tgagacgtga aaatgaatgg catgtttgga 64680 cagcattgtg tattttggtg gggtagattg taaactctgg cccaagaagt ttgagtttga 64740 tcttaaagga gccagggaaa ctgtttaagc aagggaggcc atggtaggaa atgtatatta 64800 gagtgcaaca atgtgcggag ttcactgggg gtggggagta catgctgtga ttagaaacca 64860 aaccattcgc ttccatcata ggtattttca gttaaaacaa ccttgattta aagatcataa 64920 taatgtatct ttcgtagaga tgaactttag aatttcatag agatgaatag ttggctagaa 64980 attggttgct ctattaaaat gtattttcca gataaagact ttttaagtta tttttgttga 65040 attttagaaa cttttgaaaa tcatttttaa aaaatgtacc cagatcaagg ttgcagtata 65100 aatcactgca gaacaaaatt cgtgaatctt catgtgatat ataattaaat attttggatt 65160 catccagatg aataaaatga ctaggccttt tcctcctgcc cttgttaaag gtaatgcctc 65220 tttatcaata cttcacacac acacacacac aaaaactcaa aacctttctc ttgcagttcc 65280 acggcaacct tcaaatgact tgtttgaaat atttgaaata gaaagaggag tcagtgcaga 65340 cgatgaagca aaggatgatc caggtgacac ttacactctt agtgccatca taatgacatt 65400 ttattgtttc tctccatgaa aataactttt aaatgtagat cgtttggacc atttgggaaa 65460 attacacctg ttttctttaa caactcagat tttctttgta gttaaatcag ttgagcnnnn 65520 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn tagtacttaa actgtaaaaa 65580 ggaaatcata ttttataagt gatgatgtaa cattgttata atttggcaac acttgtgtgt 65640 tctattttta cagatgatgc actattgtgc tgagaaaaga ttaatgtaat ttccctttgg 65700 tcttatggaa agaagctctg gttatatgga taggagagaa cacttgttgt ctagataccc 65760 ctagttcata gattcatttg ttgttttttg ttgattttct gatccactca tctttctgat 65820 tatttccttg taggtgttct ggtacacagt tgtaattttg accatggact ttgtggatgg 65880 atcagggaga aagacaatga cttgcactgg gaaccaatca gggacccagc aggtaaaacc 65940 atttcattta actttttctg gtacacattt caatgtgata ctatctgaag actccactgc 66000 tgctaatcaa gtctactgta acacatctct gtgtttactt gataaagatg gtcaaaaagg 66060 accatgccta gcatttatat ttcatttctt tgaaagagtt aaatttgtta gctatcaact 66120 agcactatat tatgggagac aagtagttaa ttaaaaggtc aactaatttc tctcaaaaag 66180 cttgataaat aaaatattta gattaattct tgcaaataat tcttaagtta tttgtagcac 66240 ccattcccag gaataaaaga agtaatatag tggtatatca gccatggtaa taggcattcc 66300 ccagttgnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnagat ttgagtggta 66360 gtagaagaac acagtggaaa tttctataaa gttgtgatta tcctctgaaa aaatttcact 66420 ctaacttggg tgctgaagga aatggttacg aatcaccacc actcatttta taacaaaagg 66480 tagatttaga catagttcct tggaactcaa aggagtttta aattgcacaa ttcctattaa 66540 tctaaagttt tagtactcat aagacatctt ttcttctaga ttcaagatag cttttctccc 66600 ctcagtagtt aaatcnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 66660 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 66720 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 66780 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 66840 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 66900 nnnnnnnnnn nnnnnngaag ttaaatcttt tatggataag gcctacttaa tggaagggtg 66960 aggagtttgg tttcattttt tttccttttt atactcctga ttctagattt taagaagata 67020 cagaatgtga aggaaaagct tgttatttca tttattttaa aagcctagtt cttgaatacc 67080 atactggata agagcaccaa acacagaaaa gttaacttga gagagctcat gatatcttaa 67140 accagctcnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 67200 nnnnntcatg catttctagc aggctccctg aagatctcta tggtggtcct caacgcacat 67260 tttgagtaac aaggtattaa aaaattagtn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 67320 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnatttac ttaaaaatta 67380 aaatgtgaat taaattcctt aaaataggaa tattcaataa attccaaatt ggttcataca 67440 tttcttttca ttgtaaaaga caatatcatt tagtgcctca cacacagtac tctcagtcat 67500 tcttgcatta ctgtcttact catttaaagt taactctgag caggacagca ttatcattaa 67560 tgtcacagat tctgtgaata agagagaaac caacatatct acnnnnnnnn nnnnnnnnnn 67620 nnnnntacaa tataaaatac ctatgacagt cctagcttat ttaaaatctc atcaatttta 67680 tttaattgta gcaattaaaa taaatttatt tcattgctac agagttgtag caatttacac 67740 tcctaccaac agtatttgaa aaccatcccc aacagatatt ataaaagtat ttttttaaat 67800 ctctaccaat cttataggtg aaaaggaatc tcttttaatt ttactttgca tttcttctat 67860 tgtgaaaagt tgatcatgtt ttatcattca ctcgaggagc aaannnnnnn nnnnnnnnnn 67920 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 67980 nnnnnnnnnn nnnnnnnnnn nnnnctctgt actactcatg caggctccta gacagtccaa 68040 tactcagaac atctgcagca ttattgtgtt tttgactgta ccacaggctg gcctcagtag 68100 gtgtgcatgt ggtagtgtgc atgatctgat tagcagatca cagacctgta gattagactg 68160 actgaccact gacctggagt agtttgctac agaaggaggg atctttggca gtcatgtatg 68220 acagaatggt gaataatatt tatggtcata atgaatgatt ccttcaagta agttacactg 68280 gggagagtaa ttcatgctaa actcaggaga attcactgca cttctatgta aacaatctat 68340 gtaaaatact tgagttttag aatttaaata ctgtatttta gaaccaattt agtccttttc 68400 aacatttttt tattattcat tatatttgta aatgtttaaa tttgttgtga cccaaatctc 68460 cctttctctc tgatggaaaa atgagaagag ggttagaaat tagaatggta gacaaagaaa 68520 tttgaaatcg ttattggacc gtgttagtac agagcaaatt gaaaggaaga gatgcctgct 68580 tgagtgattg gataactcgt gatgattggc tacggtccct ggaggatgct tttctagaat 68640 tggtccaagt ttttaccaag aacttagata aataggacat tgaacttaat attgaagata 68700 aactgaccaa actacggttg actcaaatct ggagaggaaa gtgactatgt gtgctgatac 68760 aaccagcatt cgaaaaatta gattaatggg tcacaattaa gaaggtgaaa tgtaatgggg 68820 ttgagtcgaa gttatggctc atagtcgaga aaccagtcac acatctgagt gagggggaag 68880 acctgacata tcactaaggg tagaagactt tagagtttta gtgtgtaatg aaatcaatct 68940 aagcgaacct tgtgatgttg ctgccagaaa tatggaccat ctctatagaa atatgctatg 69000 tttcaagaat gaaaatccac tttggttaag tggtgaatcc acttttgaga attctttttt 69060 tctaatatga gaagaataga gttggtcact tagaagagga tgacctgcat ggaaaggtat 69120 ctccaagctt gtgaaaaaca ggcaaaggaa ccataacgtt tagcctggag aagaaatgat 69180 gcatcagggt tccttttcaa ataattgact agggttagag ttgttctctg tgaccgaaag 69240 gggatgagaa taggattgat gggtaaaaaa tacagggaga ttcattttgg ctcactgtca 69300 tctaaaaaga gaacaggctg acatggaaag agtgagttat cttggtggaa gcatttgact 69360 gggtgacata tggcaagata ctgttaaaag gttttgagca tcagatgaaa gttggattag 69420 aaaacctcta aaccagctct gagagcctga ttcagaatgg aatgtggaaa tagtcccaag 69480 aaattcagtg agcaggattc tagcttggaa agaaggatgg gaagaccaaa agatggcaga 69540 gagttggaag gcaggatatg tgaggatgga attgcagaat atatttttag aatatgttga 69600 gagaattata acccagcact gatgggaaaa tactaataaa gcttaccttt tcatggtttt 69660 ttattctagg aaattcaggt ggatggannn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 69720 nnnnnnnnnn nnnnnnnnnn nnnnnnnntc acatgtgaag gccagagaag aatataaaaa 69780 atcnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 69840 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 69900 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 69960 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 70020 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 70080 aataatttct gtgtattatt ctaattacca agctaattat ttacctattt ctatctagat 70140 tataccactt ataaaagaat atgtgtttgg atatccaatt atggatgatt ttaatgatgt 70200 gcagttctaa tccaagatcc agatttgtaa atattcttat atgctaatat ttctaagaaa 70260 actctcaaaa ctcaaaacct taagaaatag ctgcaaaata agtgcatttg ctagctgcct 70320 ttcatggtgc tattaggttt tatcacattc agtcacagtg agatgtgaaa ttaacaatgc 70380 ctctaagaaa tggaaaattg tgctgctgac agacatggtg atgcgctgca ggtatgtagc 70440 tgtggccctg gggacacacc cttctcctgt gtgtgtcaaa gctagaacta aggccctttt 70500 cctggaacct tgatctctgg agaacaagga ttaagggcaa ctgactcagc actctgtctg 70560 ccactgatag ctacaaggaa tttgcttaca tatgtatcaa gtatgccagc atgtacatac 70620 aagtatgtct atttcagcat taaaacatta ttttaaaata cacataaatg taaaataaat 70680 caacaatata agtgaaagct ttctaccacc cctgactccc cattcaccac ccagcaagat 70740 gcagccagtt tcttgattat gttctcagag agatattttt ggatacaatg gcattctctt 70800 ctgtgccttg cctttgtcac ttatcgatac taccttaaga gttaacaggn nnnnnnnnnn 70860 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 70920 nnnnnnnnnn nnnnntgggg ataataagta ctatccacta nnnnnnnnnn nnnnnnnnnn 70980 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnngc tgaagcatgt 71040 cccctgtcag tacatacaga acaccnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 71100 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 71160 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 71220 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 71280 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 71340 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 71400 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 71460 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnntcat tttttctctg atgttcttta 71520 tttttcttta gtatttttat tgatttatgg aagttttcta tacaaaaaga aaaattatcc 71580 ttttttaatt tgttgcaaat atttaaaaac atttttgttt gtctatttta tggtattttt 71640 atcttaacat gcaaatttgg aaacttaaat gttcttcagt aagaaattgg ctaaatggtt 71700 attggcatac ttgcagcaat gcaaagtgta tttgctacca tagaaaaatg gttatacaag 71760 aagcaactta caacactgca aatatagtat tccttttttt aaaaaaagtt catattcatg 71820 tgcatttatg tgaatgtatt acataagaaa agttcagaag tcaaaacctt cacaaatacc 71880 tctagatttt gtgaattgaa aatttaaagg caccaaccac tacagaatta aaaataaata 71940 aactaacatt ttataaccca actttctggt caagttttgc agcatctatt atttaccctc 72000 cctccccaaa attcttgaga tattgtgata ctgtaaactg aatgaatgtt taatataatt 72060 gtacatttat caccagaatt atgaaatcag aagtgaaaat tttaataggt aacttgcttg 72120 acactgaatc aatttttccc ttagttaatg ttccaacttg tatcatttcc tttattacaa 72180 agggaacgaa ttcttcatgg nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 72240 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 72300 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 72360 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 72420 nnnnnnnnnn nnnnnnnnnn nnnnnnntac ctgtgatgat tcaaaagctg acttatagtg 72480 agacatgtaa atattttggc ccaaaatatt gtttggatta gtttgtggag gaaggggaca 72540 agggaagtgg gagaaagatt cttaagacat tatgtatttt tagtgattca agtacgtgaa 72600 tcaaggatct atttccctgc agttttttgc tgtatccttg gaacatcatt cgttgattca 72660 aacatcctta aacacctgct gcctgctagc agggtaacac agaggtttgg gggaaacttt 72720 ggttcagact tttaagttgt tcacaacctg ctggggagcc cttagcacag aggctggcac 72780 acagtaagtg cacagaaatc acctgtgacc tatagagctc cgacgggact tgttagagaa 72840 gacactggaa taaaaagtta ataccaaatt ttatcatgtc ctgtataaat taagcataag 72900 aaagagatga tagtttatat ctgttaactg aaaactaaat gggattacta ccaaaataga 72960 tacacatgtt caacattcca accatttttc tatggaaaaa ccagtgttga aacatacagt 73020 gtgttttcag ttgactgaga taaaatagcc tgagggaatg tcatttgtac atggatcaag 73080 tttcaaaata ctacttgcaa ctttctctct cttgagaagc agcacccccc acttcactgc 73140 tgcttaacca ttttttttct aatgcagttt ccttaacaac agagggaaga aaactctgta 73200 gccctaggga tctgagaaca tcatcaaagt ggtctatcta aagtgaaagt ttttattaat 73260 atttgtgttc atgtatatat aataggacat atttatagaa tagatatgat atagatttgt 73320 tttactataa aaatgtttta aatggcttac ctccaaataa aattgaagct ccaggaagac 73380 ggggagggtt tcctgaatac ctctgtatct tcctggtgtt agatttacac cagtgtgaga 73440 agctctgcca actctgattt tttttttaaa gaacctttac gtttagagat tttttttttt 73500 taagttgggg gaggagagaa taagcagata tagggccctt taaatcacca tctgctttac 73560 tgttcagagg ttaattatcc accagaaaat tctccctaga aatttagggt cagttttacc 73620 aaaccaccaa gccagatcgc caggggtgtt ggacaacgtt atcataaact ggtgagggtc 73680 acaaggtggt gaatgtgtat atgttgtcgt atgcctatat gtctgatcat attccattgg 73740 aaatattcat agtgtttggt tactaattat tgtgtgtctc tggcattact tgtactttcc 73800 atgagcaaag taagtgaatg ttttctgtaa acacatacca tagaacttaa gacaggcata 73860 caaataagtc atatttttct tcttcaaaaa tctgggaatt ctttacaaag aaaagaaaaa 73920 aaaaaaaaaa ctaaaactcc tgtccatgct gcttgttcta gaaagggata ttggcacata 73980 cacacagccc tgcttttccc attccaggtg gacaatatct gacagtgtcg gcagccaaag 74040 ccccaggggg aaaagctgca cgcttggtgc tacctctcgg ccgcctcatg cattcagggg 74100 acctgtgcct gtcattcagg cacaaggtga cggggctgca ctctggcaca ctccaggtgt 74160 ttgtgagaaa acacggtgcc cacggagcag ccctgtgggg aagaaatggt ggccatggct 74220 ggaggcaaac acagatcacc ttgcgagggg ctgacatcaa gagcgtaagt agatccacaa 74280 aggaggcagg acctgggacg ttttcctttc ataggagaac tctgggatct gaatttgaag 74340 aagccttgct gtgtgaattc aggctcagat aaaggtttgg gtttttttct gggcctgatg 74400 actccattca gtgtctcttt ctgaaaccac tttctgcttc ttcatccttc ttcattgcaa 74460 cctgtgatca ccacctctcc ctaaccctgg taaagatcca ttttttttta atggaaaaaa 74520 aaatctagga atagaaaagt ttttttggtt ttttttaaag ttgatcaaca gaaacttcca 74580 agggtggggt ggtgagctta attgttcctg ctttcccttt ccttttaatt ccttagattt 74640 caaactttta ctaccataaa ttacccagga gatggttctt ttttcttttt tgaaatgttt 74700 ctttcaagtt caaatttttt tctttgtttc agagatcaca gttaagcagc gtagggtggg 74760 aactcagaac tacaattgga aagcactatt ctattcagga caggatgtgg gaagtgcttt 74820 gatggaatat ataaatttga tcttaagtaa tcaagacata ggtagcaaaa aaatggtcta 74880 gggcatccca acaaaattat gaatgcattt gggaaaatgt catcattttg ctggtgctta 74940 tgaaacttgc tgttgggaga tacctatgcc tgtatgtagg tatagtttat aagaaaccat 75000 tttagaaaga aaagatagat tccattaata cagtttttat cagagtactt ggattttgtt 75060 taattcttac atattttttc ttaaaacttt tctcagtatt tttattgttt agagaaataa 75120 aacaagataa tcattttaaa tcatagcact tacgttttct cttgttttat aaggagcaag 75180 gatgctctat agaaaatata atgtaagaat aataaaagtt tttggttttt acataggtaa 75240 agcaacagtg tgattggatt atggtgtttg attctattcc attttcagca agaaagcgat 75300 gttaaccaca aaggaactaa gaaacattta agataggctg tgtgattatg atctttcaga 75360 tctttggctc ctaatatctg ttcctttata ttctatcaca ctcttctaac tttggtaatc 75420 cttgacaaaa gtgtgcactt tataaacaat cctaaatcga attggtctat agcttagaat 75480 ggctttttaa agaataattg attctgagta atgtggtctg atgaacagtt tgatgacttc 75540 agtttctact gaaagagaag cttcagtgat actggcaact atattctgtt tttttcctcc 75600 tgcaaaataa gtttaaaatt ggtttggggg aaggtttgcc tttatttttg cttaataagg 75660 aggcattaga aaggggcaga ggaggcttga ctggtgtgtg cattctctcc ctaggtcgtc 75720 ttcaaaggtg aaaaaaggcg tggtcacact ggggagattg gattagatga tgtgagcttg 75780 aaaaaaggcc actgctctga agaacgctaa caactccaga actaacaatg aactcctatg 75840 ttgctctatc ctctttttcc aattctcatc ttctctcctc ttctcccttt tatcaggcct 75900 aggagaagag tgggtcagtg ggtcagaagg aagtctattt ggtgacccag gtttttctgg 75960 cctgcttttg tgcaatccca atgaacagtg ataccctcct tgaaatacag gggcatcgca 76020 gacacatcaa agccatctgt gggtgttgcc ttccatcctg tgtctctttc aggaaggcat 76080 tcagcatgcg tgagccatac catcctccat cctgattaca aggtgctcct tgtagcaaat 76140 tatgagagtg agttacggga gcagttttta aaagaaatct ttgcagatgg ctatgatgtt 76200 atgtgttcgg tgttgtacca tgagtagtat tgacttccct tgagatatga tgtacaatgt 76260 gcttgtgaaa ttgacttacc ctcttcactt aagttagttc tggcctgacc tgaactctga 76320 cttttactgc cattcacttt ataaaataag ggtgtgtaac atatcaagat acatttattt 76380 ttatctgttt tttttttcct gttaaagaca attatgtaga gtgggcacgt aatccctcct 76440 tagtagtatt gtgttttgtg taaatgtgct attgatatta agtatttaca tgttccaaat 76500 atttacagac tctagttgca aggtaaaggg cagcttgtga tctcaaaaaa atacatggtg 76560 aaatgtcatc cagttccatg accttatatt ggcagcagta ggaaattggc agaagtgttg 76620 ggttgtggta acggagtgat gaattttttt ttaatggcct tgagtttgat ctctgcaaag 76680 gataggaaac ctttaggaag acaagaaact gcagttaatt tagaactgtc actgtttcaa 76740 gttacacttt aaaaccacag cttttaccat cataacatgg ctctggtaat atgtaggaag 76800 ctttataaaa gttttggttg attcagaaaa aggatcctgt tgcagagtga gaggaagcat 76860 agggggaaac tccattggaa cagattttca cacaacgttt taaattgata taagtttagg 76920 cagttgtagt tcataactta tgttgctcat gttgtgctgt gtcaggatgg gataggaagc 76980 aagtcccatg cttagaggca tgggatgtgt tggaacggga tttacacaca ctggaggagc 77040 agggcaagtt ggaattctaa gatccatgaa cccccaactg tatttcctcc ctgcatattt 77100 taccaatata ttaaaaaaca atgtaacttt taaaaggcat cattcctgag gtttgtctta 77160 atttctgatt aagtaatcag aatattttct gctgtttttg ccaggaatca caaagatgat 77220 taaagggttg gaaaaaaaga tctatgatgg aaaattaaag gaactgggat tattgagcct 77280 ggagaagaga agactgaggg gcaaaccatt gatggttttc aagtatatga agggttggca 77340 cagagagggt ggcgaccagc tgttctccat atgcactaag aatagaacaa gaggaaactg 77400 gcttagacta gagtataagg gagcatttct tggcaggggc cattgttaga atacttcata 77460 aaaaaagaag tgtgaaaatc tcagtatctc tctctctttc taaaaaatta gataaaaatt 77520 tgtctattta agatggttaa agatgttctt acccaaggaa aagtaacaaa ttatagaatt 77580 tcccaaaaga tgttttgatc ctactagtag tatgcagtga aaatctttag aactaaataa 77640 tttggacaag gcttaattta ggcatttccc tcttgacctc ctaatggaga gggattgaaa 77700 ggggaagagc ccaccaaatg ctgagctcac tgaaatatct ctcccttatg gcaatcctag 77760 cagtattaaa gaaaaaagga aactatttat tccaaatgag agtatgatgg acagatattt 77820 tagtatctca gtaatgtcct agtgtggcgg tggttttcaa tgtttcttca tggtaaaggt 77880 ataagccttt catttgttca atggatgatg tttcagattt tttttttttt aagagatcct 77940 tcaaggaaca cagttcagag agattttcat cgggtgcatt ctctctgctt cgtgtgtgac 78000 aagttatctt ggctgctgag aaagagtgcc ctgccccaca ccggcagacc tttccttcac 78060 ctcatcagta tgattcagtt tctcttatca attggactct cccaggttcc acagaacagt 78120 aatatttttt gaacaatagg tacaatagaa ggtcttctgt catttaacct ggtaaaggca 78180 gggctggagg gggaaaataa atcattaagc ctttgagtaa cggcagaata tatggctgta 78240 gatccatttt taatggttca tttcctttat ggtcatataa ctgcacagct gaagatgaaa 78300 ggggaaaata aatgaaaatt ttacttttcg atgccaatga tacattgcac taaactgatg 78360 gaagaagtta tccaaagtac tgtataacat cttgtttatt atttaatgtt ttctaaaata 78420 aaaaatgtta gtggttttcc aaatggccta ataaaaacaa ttatttgtaa ataaaaacac 78480 tgttagtaat accagttgtc tattcttgtt ttttgagttt tgtttttttt tgacttggaa 78540 aaaagcattg aggtagttaa atgatgtttc acaaaagtca tagtagaatc ccttttactg 78600 tttggatggt gggaacaaag atgttgcctg cagtattata ctttctaggt tataaaacat 78660 gagacacttt atttttttta tcagcatgaa cagggaaaga gatcagaaga tcactataac 78720 ccatgccatg ccttagtaaa ttgctttagt tatgttttat tatcatttca ttgtaaacat 78780 ttgct 78785 4 609 PRT Mus musculus 4 Met Ala Val Leu Leu Ala Ala Val Leu Ala Ser Ser Leu Tyr Leu Gln 1 5 10 15 Val Ala Ala Asp Phe Asp Gly Arg Trp Pro Arg Gln Ile Val Ser Ser 20 25 30 Ile Gly Leu Cys Arg Tyr Gly Gly Arg Ile Asp Cys Cys Trp Gly Trp 35 40 45 Ala Arg Gln Ser Trp Gly Gln Cys Gln Pro Phe Tyr Val Leu Arg Gln 50 55 60 Arg Leu Ala Arg Ile Arg Cys Gln Leu Lys Ala Val Cys Gln Pro Gln 65 70 75 80 Cys Lys His Gly Glu Cys Val Gly Pro Asn Lys Cys Lys Cys His Pro 85 90 95 Gly Phe Ala Gly Lys Thr Cys Asn Gln Asp Glu Ser Phe His Pro Thr 100 105 110 Pro Leu Asp Gln Gly Ser Glu Gln Pro Leu Phe Gln Pro Pro Asp His 115 120 125 Gln Ala Thr Asn Val Pro Ser Arg Asp Leu Asn Glu Cys Gly Leu Lys 130 135 140 Pro Arg Pro Cys Lys His Arg Cys Met Asn Thr Phe Gly Ser Tyr Lys 145 150 155 160 Cys Tyr Cys Leu Asn Gly Tyr Met Leu Leu Pro Asp Gly Ser Cys Ser 165 170 175 Ser Ala Leu Ser Cys Ser Met Ala Asn Cys Gln Tyr Gly Cys Asp Val 180 185 190 Val Lys Gly Gln Val Arg Cys Gln Cys Pro Ser Pro Gly Leu Gln Leu 195 200 205 Ala Pro Asp Gly Arg Thr Cys Val Asp Ile Asp Glu Cys Ala Thr Gly 210 215 220 Arg Val Ser Cys Pro Arg Phe Arg Gln Cys Val Asn Thr Phe Gly Ser 225 230 235 240 Tyr Ile Cys Lys Cys His Thr Gly Phe Asp Leu Met Tyr Ile Gly Gly 245 250 255 Lys Tyr Gln Cys His Asp Ile Asp Glu Cys Ser Leu Gly Gln His Gln 260 265 270 Cys Ser Ser Tyr Ala Arg Cys Tyr Asn Ile His Gly Ser Tyr Lys Cys 275 280 285 Gln Cys Arg Asp Gly Tyr Glu Gly Asp Gly Leu Asn Cys Val Tyr Ile 290 295 300 Pro Lys Val Met Ile Glu Pro Ser Gly Pro Ile His Met Pro Glu Arg 305 310 315 320 Asn Gly Thr Ile Ser Lys Gly Asp Gly Gly His Ala Asn Arg Ile Pro 325 330 335 Asp Ala Gly Ser Thr Arg Trp Pro Leu Lys Thr Pro Tyr Ile Pro Pro 340 345 350 Val Ile Thr Asn Arg Pro Thr Ser Lys Pro Thr Thr Arg Pro Thr Pro 355 360 365 Asn Pro Thr Pro Gln Pro Thr Pro Pro Pro Pro Pro Pro Leu Pro Thr 370 375 380 Glu Pro Arg Thr Thr Pro Leu Pro Pro Thr Pro Glu Arg Pro Ser Thr 385 390 395 400 Arg Pro Thr Thr Ile Ala Pro Ala Thr Ser Thr Thr Thr Arg Val Ile 405 410 415 Thr Val Asp Asn Arg Ile Gln Thr Asp Pro Gln Lys Pro Arg Gly Asp 420 425 430 Val Phe Ile Pro Arg Gln Pro Thr Asn Asp Leu Phe Glu Ile Phe Glu 435 440 445 Ile Glu Arg Gly Val Ser Ala Asp Glu Glu Val Lys Asp Asp Pro Gly 450 455 460 Ile Leu Ile His Ser Cys Asn Phe Asp His Gly Leu Cys Gly Trp Ile 465 470 475 480 Arg Glu Lys Asp Ser Asp Leu His Trp Glu Thr Ala Arg Asp Pro Ala 485 490 495 Gly Gly Gln Tyr Leu Thr Val Ser Ala Ala Lys Ala Pro Gly Gly Lys 500 505 510 Ala Ala Arg Leu Val Leu Arg Leu Gly His Leu Met His Ser Gly Asp 515 520 525 Leu Cys Leu Ser Phe Arg His Lys Val Thr Gly Leu His Ser Gly Thr 530 535 540 Leu Gln Val Phe Val Arg Lys His Gly Thr His Gly Ala Ala Leu Trp 545 550 555 560 Gly Arg Asn Gly Gly His Gly Trp Arg Gln Thr Gln Ile Thr Leu Arg 565 570 575 Gly Ala Asp Val Lys Ser Val Ile Phe Lys Gly Glu Lys Arg Arg Gly 580 585 590 His Thr Gly Glu Ile Gly Leu Asp Asp Val Ser Leu Lys Arg Gly Arg 595 600 605 Cys 5 592 PRT Mus musculus 5 Met Ala Val Leu Leu Ala Ala Val Leu Ala Ser Ser Leu Tyr Leu Gln 1 5 10 15 Val Ala Ala Asp Phe Asp Gly Arg Trp Pro Arg Gln Ile Val Ser Ser 20 25 30 Ile Gly Leu Cys Arg Tyr Gly Gly Arg Ile Asp Cys Cys Trp Gly Trp 35 40 45 Ala Arg Gln Ser Trp Gly Gln Cys Gln Pro Val Cys Gln Pro Gln Cys 50 55 60 Lys His Gly Glu Cys Val Gly Pro Asn Lys Cys Lys Cys His Pro Gly 65 70 75 80 Phe Ala Gly Lys Thr Cys Asn Gln Asp Glu Ser Phe His Pro Thr Pro 85 90 95 Leu Asp Gln Gly Ser Glu Gln Pro Leu Phe Gln Pro Pro Asp His Gln 100 105 110 Ala Thr Asn Val Pro Ser Arg Asp Leu Asn Glu Cys Gly Leu Lys Pro 115 120 125 Arg Pro Cys Lys His Arg Cys Met Asn Thr Phe Gly Ser Tyr Lys Cys 130 135 140 Tyr Cys Leu Asn Gly Tyr Met Leu Leu Pro Asp Gly Ser Cys Ser Ser 145 150 155 160 Ala Leu Ser Cys Ser Met Ala Asn Cys Gln Tyr Gly Cys Asp Val Val 165 170 175 Lys Gly Gln Val Arg Cys Gln Cys Pro Ser Pro Gly Leu Gln Leu Ala 180 185 190 Pro Asp Gly Arg Thr Cys Val Asp Ile Asp Glu Cys Ala Thr Gly Arg 195 200 205 Val Ser Cys Pro Arg Phe Arg Gln Cys Val Asn Thr Phe Gly Ser Tyr 210 215 220 Ile Cys Lys Cys His Thr Gly Phe Asp Leu Met Tyr Ile Gly Gly Lys 225 230 235 240 Tyr Gln Cys His Asp Ile Asp Glu Cys Ser Leu Gly Gln His Gln Cys 245 250 255 Ser Ser Tyr Ala Arg Cys Tyr Asn Ile His Gly Ser Tyr Lys Cys Gln 260 265 270 Cys Arg Asp Gly Tyr Glu Gly Asp Gly Leu Asn Cys Val Tyr Ile Pro 275 280 285 Lys Val Met Ile Glu Pro Ser Gly Pro Ile His Met Pro Glu Arg Asn 290 295 300 Gly Thr Ile Ser Lys Gly Asp Gly Gly His Ala Asn Arg Ile Pro Asp 305 310 315 320 Ala Gly Ser Thr Arg Trp Pro Leu Lys Thr Pro Tyr Ile Pro Pro Val 325 330 335 Ile Thr Asn Arg Pro Thr Ser Lys Pro Thr Thr Arg Pro Thr Pro Asn 340 345 350 Pro Thr Pro Gln Pro Thr Pro Pro Pro Pro Pro Pro Leu Pro Thr Glu 355 360 365 Pro Arg Thr Thr Pro Leu Pro Pro Thr Pro Glu Arg Pro Ser Thr Arg 370 375 380 Pro Thr Thr Ile Ala Pro Ala Thr Ser Thr Thr Thr Arg Val Ile Thr 385 390 395 400 Val Asp Asn Arg Ile Gln Thr Asp Pro Gln Lys Pro Arg Gly Asp Val 405 410 415 Phe Ile Pro Arg Gln Pro Thr Asn Asp Leu Phe Glu Ile Phe Glu Ile 420 425 430 Glu Arg Gly Val Ser Ala Asp Glu Glu Val Lys Asp Asp Pro Gly Ile 435 440 445 Leu Ile His Ser Cys Asn Phe Asp His Gly Leu Cys Gly Trp Ile Arg 450 455 460 Glu Lys Asp Ser Asp Leu His Trp Glu Thr Ala Arg Asp Pro Ala Gly 465 470 475 480 Gly Gln Tyr Leu Thr Val Ser Ala Ala Lys Ala Pro Gly Gly Lys Ala 485 490 495 Ala Arg Leu Val Leu Arg Leu Gly His Leu Met His Ser Gly Asp Leu 500 505 510 Cys Leu Ser Phe Arg His Lys Val Thr Gly Leu His Ser Gly Thr Leu 515 520 525 Gln Val Phe Val Arg Lys His Gly Thr His Gly Ala Ala Leu Trp Gly 530 535 540 Arg Asn Gly Gly His Gly Trp Arg Gln Thr Gln Ile Thr Leu Arg Gly 545 550 555 560 Ala Asp Val Lys Ser Val Ile Phe Lys Gly Glu Lys Arg Arg Gly His 565 570 575 Thr Gly Glu Ile Gly Leu Asp Asp Val Ser Leu Lys Arg Gly Arg Cys 580 585 590 6 578 PRT Mus musculus 6 Met Ala Val Leu Leu Ala Ala Val Leu Ala Ser Ser Leu Tyr Leu Gln 1 5 10 15 Val Ala Ala Asp Phe Asp Gly Arg Trp Pro Arg Gln Ile Val Ser Ser 20 25 30 Ile Gly Leu Cys Arg Tyr Gly Gly Arg Ile Asp Cys Cys Trp Gly Trp 35 40 45 Ala Arg Gln Ser Trp Gly Gln Cys Gln Pro Phe Tyr Val Leu Arg Gln 50 55 60 Arg Leu Ala Arg Ile Arg Cys Gln Leu Lys Ala Val Cys Gln Pro Gln 65 70 75 80 Cys Lys His Gly Glu Cys Val Gly Pro Asn Lys Cys Lys Cys His Pro 85 90 95 Gly Phe Ala Gly Lys Thr Cys Asn Gln Asp Leu Asn Glu Cys Gly Leu 100 105 110 Lys Pro Arg Pro Cys Lys His Arg Cys Met Asn Thr Phe Gly Ser Tyr 115 120 125 Lys Cys Tyr Cys Leu Asn Gly Tyr Met Leu Leu Pro Asp Gly Ser Cys 130 135 140 Ser Ser Ala Leu Ser Cys Ser Met Ala Asn Cys Gln Tyr Gly Cys Asp 145 150 155 160 Val Val Lys Gly Gln Val Arg Cys Gln Cys Pro Ser Pro Gly Leu Gln 165 170 175 Leu Ala Pro Asp Gly Arg Thr Cys Val Asp Ile Asp Glu Cys Ala Thr 180 185 190 Gly Arg Val Ser Cys Pro Arg Phe Arg Gln Cys Val Asn Thr Phe Gly 195 200 205 Ser Tyr Ile Cys Lys Cys His Thr Gly Phe Asp Leu Met Tyr Ile Gly 210 215 220 Gly Lys Tyr Gln Cys His Asp Ile Asp Glu Cys Ser Leu Gly Gln His 225 230 235 240 Gln Cys Ser Ser Tyr Ala Arg Cys Tyr Asn Ile His Gly Ser Tyr Lys 245 250 255 Cys Gln Cys Arg Asp Gly Tyr Glu Gly Asp Gly Leu Asn Cys Val Tyr 260 265 270 Ile Pro Lys Val Met Ile Glu Pro Ser Gly Pro Ile His Met Pro Glu 275 280 285 Arg Asn Gly Thr Ile Ser Lys Gly Asp Gly Gly His Ala Asn Arg Ile 290 295 300 Pro Asp Ala Gly Ser Thr Arg Trp Pro Leu Lys Thr Pro Tyr Ile Pro 305 310 315 320 Pro Val Ile Thr Asn Arg Pro Thr Ser Lys Pro Thr Thr Arg Pro Thr 325 330 335 Pro Asn Pro Thr Pro Gln Pro Thr Pro Pro Pro Pro Pro Pro Leu Pro 340 345 350 Thr Glu Pro Arg Thr Thr Pro Leu Pro Pro Thr Pro Glu Arg Pro Ser 355 360 365 Thr Arg Pro Thr Thr Ile Ala Pro Ala Thr Ser Thr Thr Thr Arg Val 370 375 380 Ile Thr Val Asp Asn Arg Ile Gln Thr Asp Pro Gln Lys Pro Arg Gly 385 390 395 400 Asp Val Phe Ile Pro Arg Gln Pro Thr Asn Asp Leu Phe Glu Ile Phe 405 410 415 Glu Ile Glu Arg Gly Val Ser Ala Asp Glu Glu Val Lys Asp Asp Pro 420 425 430 Gly Ile Leu Ile His Ser Cys Asn Phe Asp His Gly Leu Cys Gly Trp 435 440 445 Ile Arg Glu Lys Asp Ser Asp Leu His Trp Glu Thr Ala Arg Asp Pro 450 455 460 Ala Gly Gly Gln Tyr Leu Thr Val Ser Ala Ala Lys Ala Pro Gly Gly 465 470 475 480 Lys Ala Ala Arg Leu Val Leu Arg Leu Gly His Leu Met His Ser Gly 485 490 495 Asp Leu Cys Leu Ser Phe Arg His Lys Val Thr Gly Leu His Ser Gly 500 505 510 Thr Leu Gln Val Phe Val Arg Lys His Gly Thr His Gly Ala Ala Leu 515 520 525 Trp Gly Arg Asn Gly Gly His Gly Trp Arg Gln Thr Gln Ile Thr Leu 530 535 540 Arg Gly Ala Asp Val Lys Ser Val Ile Phe Lys Gly Glu Lys Arg Arg 545 550 555 560 Gly His Thr Gly Glu Ile Gly Leu Asp Asp Val Ser Leu Lys Arg Gly 565 570 575 Arg Cys 

That which is claimed is:
 1. An isolated peptide consisting of an amino acid sequence selected from the group consisting of: (a) an amino acid sequence shown in SEQ ID NO:2; (b) an amino acid sequence of an allelic variant of an amino acid sequence shown in SEQ ID NO:2, wherein said allelic variant is encoded by a nucleic acid molecule that hybridizes under stringent conditions to the opposite strand of a nucleic acid molecule shown in SEQ ID NOS:1 or 3; (c) an amino acid sequence of an ortholog of an amino acid sequence shown in SEQ ID NO:2, wherein said ortholog is encoded by a nucleic acid molecule that hybridizes under stringent conditions to the opposite strand of a nucleic acid molecule shown in SEQ ID NOS:1 or 3; and (d) a fragment of an amino acid sequence shown in SEQ ID NO:2, wherein said fragment comprises at least 10 contiguous amino acids.
 2. An isolated peptide comprising an amino acid sequence selected from the group consisting of: (a) an amino acid sequence shown in SEQ ID NO:2; (b) an amino acid sequence of an allelic variant of an amino acid sequence shown in SEQ ID NO:2, wherein said allelic variant is encoded by a nucleic acid molecule that hybridizes under stringent conditions to the opposite strand of a nucleic acid molecule shown in SEQ ID NOS:1 or 3; (c) an amino acid sequence of an ortholog of an amino acid sequence shown in SEQ ID NO:2, wherein said ortholog is encoded by a nucleic acid molecule that hybridizes under stringent conditions to the opposite strand of a nucleic acid molecule shown in SEQ ID NOS:1 or 3; and (d) a fragment of an amino acid sequence shown in SEQ ID NO:2, wherein said fragment comprises at least 10 contiguous amino acids.
 3. An isolated antibody that selectively binds to a peptide of claim
 2. 4. An isolated nucleic acid molecule consisting of a nucleotide sequence selected from the group consisting of: (a) a nucleotide sequence that encodes an amino acid sequence shown in SEQ ID NO:2; (b) a nucleotide sequence that encodes of an allelic variant of an amino acid sequence shown in SEQ ID NO:2, wherein said nucleotide sequence hybridizes under stringent conditions to the opposite strand of a nucleic acid molecule shown in SEQ ID NOS:1 or 3; (c) a nucleotide sequence that encodes an ortholog of an amino acid sequence shown in SEQ ID NO:2, wherein said nucleotide sequence hybridizes under stringent conditions to the opposite strand of a nucleic acid molecule shown in SEQ ID NOS:1 or 3; (d) a nucleotide sequence that encodes a fragment of an amino acid sequence shown in SEQ ID NO:2, wherein said fragment comprises at least 10 contiguous amino acids; and (e) a nucleotide sequence that is the complement of a nucleotide sequence of (a)-(d).
 5. An isolated nucleic acid molecule comprising a nucleotide sequence selected from the group consisting of: (a) a nucleotide sequence that encodes an amino acid sequence shown in SEQ ID NO:2; (b) a nucleotide sequence that encodes of an allelic variant of an amino acid sequence shown in SEQ ID NO:2, wherein said nucleotide sequence hybridizes under stringent conditions to the opposite strand of a nucleic acid molecule shown in SEQ ID NOS:1 or 3; (c) a nucleotide sequence that encodes an ortholog of an amino acid sequence shown in SEQ ID NO:2, wherein said nucleotide sequence hybridizes under stringent conditions to the opposite strand of a nucleic acid molecule shown in SEQ ID NOS:1 or 3; (d) a nucleotide sequence that encodes a fragment of an amino acid sequence shown in SEQ ID NO:2, wherein said fragment comprises at least 10 contiguous amino acids; and (e) a nucleotide sequence that is the complement of a nucleotide sequence of (a)-(d).
 6. A gene chip comprising a nucleic acid molecule of claim
 5. 7. A transgenic non-human animal comprising a nucleic acid molecule of claim
 5. 8. A nucleic acid vector comprising a nucleic acid molecule of claim
 5. 9. A host cell containing the vector of claim
 8. 10. A method for producing any of the peptides of claim 1 comprising introducing a nucleotide sequence encoding any of the amino acid sequences in (a)-(d) into a host cell, and culturing the host cell under conditions in which the peptides are expressed from the nucleotide sequence.
 11. A method for producing any of the peptides of claim 2 comprising introducing a nucleotide sequence encoding any of the amino acid sequences in (a)-(d) into a host cell, and culturing the host cell under conditions in which the peptides are expressed from the nucleotide sequence.
 12. A method for detecting the presence of any of the peptides of claim 2 in a sample, said method comprising contacting said sample with a detection agent that specifically allows detection of the presence of the peptide in the sample and then detecting the presence of the peptide.
 13. A method for detecting the presence of a nucleic acid molecule of claim 5 in a sample, said method comprising contacting the sample with an oligonucleotide that hybridizes to said nucleic acid molecule under stringent conditions and determining whether the oligonucleotide binds to said nucleic acid molecule in the sample.
 14. A method for identifying a modulator of a peptide of claim 2, said method comprising contacting said peptide with an agent and determining if said agent has modulated the function or activity of said peptide.
 15. The method of claim 14, wherein said agent is administered to a host cell comprising an expression vector that expresses said peptide.
 16. A method for identifying an agent that binds to any of the peptides of claim 2, said method comprising contacting the peptide with an agent and assaying the contacted mixture to determine whether a complex is formed with the agent bound to the peptide.
 17. A pharmaceutical composition comprising an agent identified by the method of claim 16 and a pharmaceutically acceptable carrier therefor.
 18. A method for treating a disease or condition mediated by a human secreted protein, said method comprising administering to a patient a pharmaceutically effective amount of an agent identified by the method of claim
 16. 19. A method for identifying a modulator of the expression of a peptide of claim 2, said method comprising contacting a cell expressing said peptide with an agent, and determining if said agent has modulated the expression of said peptide.
 20. An isolated human secreted peptide having an amino acid sequence that shares at least 70% homology with an amino acid sequence shown in SEQ ID NO:2.
 21. A peptide according to claim 20 that shares at least 90 percent homology with an amino acid sequence shown in SEQ ID NO:2.
 22. An isolated nucleic acid molecule encoding a human secreted peptide, said nucleic acid molecule sharing at least 80 percent homology with a nucleic acid molecule shown in SEQ ID NOS:1 or
 3. 23. A nucleic acid molecule according to claim 22 that shares at least 90 percent homology with a nucleic acid molecule shown in SEQ ID NOS:1 or
 3. 